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Небесная энциклопедия

Космические корабли и станции, автоматические КА и методы их проектирования, бортовые комплексы управления, системы и средства жизнеобеспечения, особенности технологии производства ракетно-космических систем

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Мониторинг СМИ

Мониторинг СМИ и социальных сетей. Сканирование интернета, новостных сайтов, специализированных контентных площадок на базе мессенджеров. Гибкие настройки фильтров и первоначальных источников.

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Форма поиска

Поддерживает ввод нескольких поисковых фраз (по одной на строку). При поиске обеспечивает поддержку морфологии русского и английского языка
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Применить Всего найдено 9999. Отображено 100.
03-03-2021 дата публикации

Устройство для получения коллагена из кожевенного сырья.

Номер: RU0000202701U1

Полезная модель относится к оборудованию для жидкостной обработки отходов шкур животных, образующихся в кожевенном производстве.Технический результат достигается тем, что внутренняя емкость устройства, в которую помещают исходное сырье для обработки, дополнительно оборудована прессом для проведения операции отжима обработанного сырья.Устройство работает следующим образом:Во внутреннюю емкость, помещают кожевенное сырье (гольевую обрезь), а во внешнюю емкость, через нижний трубопровод подают воду для проведения операции промывки кожевенного сырья, обработанное сырье отжимают прессом для удаления избытка воды и реагентов. Обработанное сырье подвергают гидротермообработке в 2-3% растворе пероксида водорода. Использование пресса для отжима сырья позволяет более точно выдержать временные ограничения проведения операции. Далее в емкость подают раствор щелочи для разволокнения молекул коллагена.Предлагаемая полезная модель позволяет быстрее и значительно эффективнее получить коллаген, требуемого качества. (Илл. 1.). РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 202 701 U1 (51) МПК C14C 15/00 (2006.01) C07K 1/36 (2006.01) C07K 14/78 (2006.01) B01F 7/00 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ПОЛЕЗНОЙ МОДЕЛИ К ПАТЕНТУ (52) СПК C14C 15/00 (2021.01); C07K 1/36 (2021.01); B01F 7/00 (2021.01) (21)(22) Заявка: 2020135782, 30.10.2020 (24) Дата начала отсчета срока действия патента: Дата регистрации: 03.03.2021 R U 2 0 2 7 0 1 U 1 Адрес для переписки: 305041, Курская обл., г. Курск, ул. Карла Маркса, 3, Лазаренко Виктор Анатольевич (56) Список документов, цитированных в отчете о поиске: RU 2066351 C1, 10.09.1996. RU 2609635 C1, 02.02.2017. RU 2049110 C1, 27.11.1995. EP 3440228 B1, 13.05.2020. U 1 (45) Опубликовано: 03.03.2021 Бюл. № 7 (73) Патентообладатель(и): федеральное государственное бюджетное образовательное учреждение высшего образования "Курский государственный медицинский университет" Министерства здравоохранения Российской Федерации (RU) 2 0 ...

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01-03-2012 дата публикации

Method for preparing protein, dna, and rna from cell

Номер: US20120053329A1
Автор: Nobuko Yamamoto
Принадлежит: Canon Inc

An object of the present invention is to prepare DNA, RNA, and a protein from one cell and to provide a convenient preparation method with high reproducibility. To prepare a protein, DNA, and RNA from a cell, nuclear and cytoplasmic separation is performed, and a protein, DNA, and RNA are then extracted.

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29-03-2012 дата публикации

Method for removing viruses from high concentration monoclonal antibody solution

Номер: US20120077963A1
Принадлежит: Asahi Kasei Medical Co Ltd

An object of the present invention is to provide a method for removing even small viruses from a high concentration monoclonal antibody solution using a membrane, and thus for recovering the antibody within a short time at high yield in the form of a filtrate. The present invention provides a method for producing a preparation containing a monoclonal antibody, which comprises a step of removing viruses by filtering viruses in a monoclonal antibody solution using a virus-removing membrane, wherein (1) the monomer content of the monoclonal antibody accounts for 90% or more; (2) the monoclonal antibody concentration in the monoclonal antibody solution ranges from 20 mg/ml to 100 mg/ml; (3) the monoclonal antibody solution contains at least a basic amino acid; and (4) the parvovirus removal rate of the virus-removing membrane satisfies the following conditions: LRV (Log Reduction Value: logarithmic reduction value) ≧4.

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31-05-2012 дата публикации

Protein crystallization using molecularly imprinted polymers

Номер: US20120135442A1
Принадлежит: UNIVERSITY OF SURREY

The invention relates to a method comprising: providing a molecularly imprinted polymer imprinted with a first peptide or protein; exposing said molecularly imprinted polymer to a supersaturated solution of a second peptide or protein; and forming a nucleus of and/or growing a crystal of said second peptide or protein on said molecularly imprinted polymer.

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24-01-2013 дата публикации

Crystalline form of cyclosporine a, methods of preparation, and methods for use thereof

Номер: US20130023482A1
Принадлежит: Allergan Inc

The present invention relates generally to crystalline forms of cyclosporine A and particularly to a newly identified form of cyclosporine A. The invention further relates to methods for its preparation and to methods for treating certain ocular disorders.

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07-02-2013 дата публикации

Method for Removing Endotoxin from Proteins

Номер: US20130035477A1
Принадлежит: Individual

Disclosed is a method for removing endotoxin from proteins. Also disclosed are products made by using the method. The method may be used, for example, to produce endotoxin-free lactoferrin. Bovine milk-derived lactoferrin may be produced in commercial quantities by the method, and endotoxin-free bovine lactoferrin may be used for a variety of therapeutic uses, including improving wound healing.

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21-02-2013 дата публикации

Antibody Preparations

Номер: US20130045199A1
Принадлежит: BIOTEST AG

An antibody preparation suitable for intravenous administration in humans includes IgG, IgA and at least 5% IgM antibodies by weight of the total amount of antibodies. The preparation is prepared from human plasma, has specific complement activating activity, and, in an in vitro assay with human serum suitable to determine the ability of the antibody preparation to activate complement unspecifically, the antibody preparation generates substantially no C5 a and/or substantially no C3 a . The antibody preparation can have medical uses.

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28-02-2013 дата публикации

Protein A Crystals and Cross-Linked Crystals and Methods of Use Thereof

Номер: US20130053548A1
Принадлежит: Individual

Protein A crystals and Protein A cross-linked protein crystals (CLPCs) are described. Methods of preparing and using are also disclosed.

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07-03-2013 дата публикации

Negatively charged porous medium for removing protein aggregates

Номер: US20130056415A1
Принадлежит: EMD Millipore Corp

A negatively charged microporous filtration medium having a high charge density comprising a porous substrate and a polymerized cross-linked polymeric coating located on the inner and outer surfaces of the substrate. The coating may be formed from a reactant solution comprising negatively charged cross-linkable polymerizeable acrylamidoalkyl monomers and acrylamido cross-linking agents which are polymerized in situ on the substrate. The negatively charged microporous filtration medium are suitable for use as prefiltration membranes for selectively removing protein aggregates from a protein solution.

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07-03-2013 дата публикации

Crystalline d-isoglutamyl-d-tryptophan and the mono ammonium slat of d-isoglutamyl-d-tryptophan

Номер: US20130059791A1
Принадлежит: Apotex Technologies Inc

A process for making pure crystalline D-isoglutamyl-D-tryptophan is provided which includes the step of deprotecting essentially pure N-tert-butoxycarbonyl-D-isoglutamyl-D-tryptophan or its diester to yield essentially pure D-isoglutamyl-D-tryptophan. A process is also provided for the preparation of pure mono ammonium salt of D-isoglutamyl-D-tryptophan from essentially pure N-tert-butoxycarbonyl-D-isoglutamyl-D-tryptophan. D-isoglutamyl-D-tryptophan, ammonium salt (1:1) is a stable pharmaceutical solid.

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07-03-2013 дата публикации

Crystalline d-isoglutamyl-d-tryptophan and the mono ammonium slat of d-isoglutamyl-d-tryptophan

Номер: US20130060048A1
Принадлежит: Apotex Technologies Inc

A process for making pure crystalline D-isoglutamyl-D-tryptophan is provided which includes the step of deprotecting essentially pure N-tert-butoxycarbonyl-D-isoglutamyl-D-tryptophan or its diester to yield essentially pure D-isoglutamyl-D-tryptophan. A process is also provided for the preparation of pure mono ammonium salt of D-isoglutamyl-D-tryptophan from essentially pure N-tert-butoxycarbonyl-D-isoglutamyl-D-tryptophan. D-isoglutamyl-D-tryptophan, ammonium salt ( 1:1 ) is a stable pharmaceutical solid.

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18-04-2013 дата публикации

VANCOMYCIN B HYDROCHLORIDE CRYSTALLINE FORM 1

Номер: US20130096275A1
Принадлежит: ABBOTT LABORATORIES

Vancomycin B Hydrochloride Crystalline Form 1, compositions containing it and methods of prevention or treatment of bacterial infections using it are disclosed. 1providing a mixture comprising Vancomycin B hydrochloride and one volume of water, wherein the vancomycin B hydrochloride is completely soluble in the water;causing Vancomycin B Hydrochloride Crystalline Form 1 to exist in the mixture by adding 1.5 volumes of acetone to the mixture; andisolating the Vancomycin B Hydrochloride Crystalline Form 1.. A process for making Vancomycin B Hydrochloride Crystalline Form 1 comprising: This application claims priority to U.S. patent application Ser. No. 60/882,656, filed Dec. 29, 2006 and is incorporated herein by reference in its entirety.This invention pertains to Vancomycin B Hydrochloride Crystalline Form 1, ways to make it, compositions containing it and methods of treatment of diseases using it.Vancomycin B hydrochloride is a salt of Vancomycin B (CHClNO) that is produced by the actinomycete Amycolaptosis orientalis (formerly Streptomyces orientalis) found in Indonesian and Indian soil.Because the relationship between different crystalline forms of Vancomycin B Hydrochloride may provide guidance for further development, there is an existing need in the chemical and therapeutic arts for identification of different crystalline forms of Vancomycin B Hydrochloride and ways of reproducibly making it.One embodiment of this invention pertains to Vancomycin B Hydrochloride Crystalline Form 1.Still another embodiment of this invention pertains to Vancomycin B Hydrochloride Crystalline Form 1 having substantial crystalline purity.Still another embodiment pertains to compositions made with or comprising an excipient and Vancomycin B Hydrochloride Crystalline Form 1.Still another embodiment pertains to methods for treating bacterial infection in a human comprising administering thereto a therapeutically effective amount of Vancomycin B Hydrochloride Crystalline Form 1.Still ...

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18-04-2013 дата публикации

PROCESS FOR THE PURIFICATION OF A GROWTH FACTOR PROTEIN

Номер: US20130096279A1
Принадлежит: Octapharma AG

A process of purifying a Growth Factor Protein in a purification sequence employing chromatography characterized in that 1. A process of purifying a Growth Factor Protein selected from the group consisting of Colony Stimulating Factor (CSF) , interleukin 3 (IL-3) , Hepatocyte growth factor , Epidermal growth factor and fibroblast growth factor (acid) in a purification sequence employing chromatography comprisingbinding the Growth Factor Protein to a multimodal resin at a pH between 4 to 6.2, andeluting the Growth Factor Protein at a pH>6.3.2. The process of wherein the multimodal resin comprises moieties bound to a matrix and the moieties are able to interact with the Growth Factor Protein in a mixture by ionic interactions claim 1 , hydrogen bonding claim 1 , and/or hydrophobic interaction.3. The process of wherein the mixture comprising the Growth Factor Protein is a solution.4. The process of wherein the Growth Factor Protein is a recombinant Growth Factor Protein.5. The process of wherein the Growth Factor Protein is eluted by a pH change>pH 6.3.6. The process of wherein the elution is performed with a change of the pH or with an elution agent comprising an amino acid having a basic side chain and/or high ionic strength in an elution buffer.7. The process according to wherein the concentration of the elution agent is in the range of from about 0.1 M to about 2.0 M.8. The process of wherein the Growth Factor Protein binds to the multimodal resin at pH about 4.0 to pH about 6.0 claim 1 , and the Growth Factor Protein is eluted from the multimodal resin at pH 6.5 or higher.9. The process of comprising using a buffering substance comprising sodium citrate claim 1 , sodium acetate claim 1 , or HEPES.10. The process of wherein a non-ionic detergent is present in any of the buffers used.11. The process of wherein the amino acid having a basic side chain is arginine claim 8 , lysine and/or histidine.12. The process of wherein buffers comprising about 0.1 M-2 M sodium ...

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18-04-2013 дата публикации

METHOD FOR PURIFYING PROTEIN USING AMINO ACID

Номер: US20130096284A1
Автор: ISHIHARA Takashi
Принадлежит: KYOWA HAKKO KIRIN CO., LTD.

In large-scale purification of proteins such as antibodies, an economic high-purity purification method is required. The present invention relates to a method for purifying a protein, including one or more chromatographic processes, in which an amino acid; or a dipeptide, an oligopeptide, or a polyamino acid thereof is included in a buffer solution used in at least one chromatographic processes (equilibration buffer, wash buffer, and elution buffer), thereby purifying a high-purity protein with a very small quantity of the impurity (e.g., polymers or host cell proteins). 1. A method for purifying a protein , comprising one or more chromatographic processes , wherein an amino acid is included as an ingredient of a buffer solution or a part thereof used in at least one chromatographic process.2. The purification method according to claim 1 , wherein an amino acid is included as an ingredient of a buffer solution or a part thereof used in a cation exchange chromatographic process.3. The purification method according to claim 2 , which further comprises claim 2 , as the chromatographic process claim 2 , a Protein A affinity chromatographic process and one or more chromatographic processes selected from an anion exchange chromatographic process claim 2 , a gel filtration chromatographic process claim 2 , a hydrophobic chromatographic process claim 2 , a hydroxyapatite chromatographic process claim 2 , and a mixed mode chromatographic process.4. The purification method according to claim 2 , wherein the cation exchange chromatographic process is carried out subsequent to the Protein A affinity chromatographic process.5. The purification method according to claim 4 , wherein the anion exchange chromatographic process is further subsequently carried out.6. The purification method according to claim 3 , wherein the cation exchange chromatographic process carried out subsequent to the Protein A affinity chromatographic process and the anion exchange chromatographic process.7. ...

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25-04-2013 дата публикации

METHOD TO PRODUCE AN IMMUNOGLOBULIN PREPARATION WITH IMPROVED YIELD

Номер: US20130101579A1
Принадлежит:

The present invention provides improved methods for the manufacturing of IVIG products. These methods offer various advantages such as reduced loss of IgG during purification and improved quality of final products. In other aspects, the present invention provides aqueous and pharmaceutical compositions suitable for intravenous, subcutaneous, and/or intramuscular administration. In yet other embodiments, the present invention provides methods of treating a disease or condition comprising administration of an IgG composition provided herein. 1. A method for preparing an enriched IgG composition from plasma , the method comprising the steps of:(a) precipitating a cryo-poor plasma fraction, in a first precipitation step, with from about 6% to about 10% alcohol at a pH of from about 7.0 to about 7.5 to obtain a first precipitate and a first supernatant;(b) precipitating IgG from the first supernatant, in a second precipitation step, with from about 23% to about 27% alcohol at a pH of from about 6.7 to about 7.1 at a temperature of from about −7° C., to about −9° C. to form a second precipitate;(c) suspending the second precipitate to form a suspension;(d) precipitating IgG from the suspension formed in step (c), in a third precipitation step, with from about 22% to about 28% alcohol at a pH of from about 6.7 to about 7.3 to form a third precipitate;(e) re-suspending the third precipitate to form a suspension; and(f) separating the soluble fraction from the suspension formed in step (e), thereby forming an enriched IgG composition.258-. (canceled)59. The method of claim 1 , wherein the temperature of precipitation step (b) is about −7° C.60. The method of claim 1 , wherein the alcohol concentration of precipitation step (b) is about 25%.61. The method of claim 59 , wherein the alcohol concentration of precipitation step (b) is about 25%.62. The method of claim 1 , wherein at least one of the first precipitation step claim 1 , second precipitation step claim 1 , or third ...

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25-04-2013 дата публикации

Particle matrix for storage of biomolecules

Номер: US20130101981A1
Принадлежит: Individual

Matrices for manipulation of biopolymers, including the separation, purification, immobilization and archival storage of biopolymers is disclosed.

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25-04-2013 дата публикации

MANUFACTURE OF FACTOR H (FH) AND FH-DERIVATIVES FROM PLASMA

Номер: US20130102537A1
Принадлежит:

The present invention provides compositions and pharmaceutical formulations of Factor H derived from plasma. Also provided are methods for the manufacture of the Factor H compositions and formulations, as well as methods for the treatment of diseases associated with Factor H dysfunction. 114-. (canceled)15. A method for preparing an enriched Factor H composition from plasma , the method comprising the steps of:(a) extracting Factor H from a Fraction I precipitate and/or a Fraction II+III precipitate, to form a Factor H extract; and(b) precipitating an impurity from the Factor H extract to form a supernatant containing Factor H, thereby preparing an enriched Factor H composition.1627-. (canceled)28. The method of claim 15 , wherein extraction step (a) comprises re-suspending the Fraction II+III precipitate with a re-suspension buffer having a pH of from 4.0 to 5.5.29. The method of claim 15 , wherein precipitation step (b) comprises precipitating an impurity from the Factor extract using a final alcohol concentration of from 10% to 20% at a pH of from 7.0 to 9.0.30. The method of claim 15 , wherein precipitation step (b) comprises precipitating an impurity from the Factor H extract using a final alcohol concentration of from 14% to 16% at a pH of from 7.5 to 8.5.31. The method of claim 15 , wherein the method further comprises the step of:(c) precipitating Factor H from the supernatant formed in step (h) using a final alcohol concentration of from 20% to 30% at a pH of from 5.0 to 7.0.32. The method of claim 15 , wherein the method further comprises the step of:(c) precipitating Factor H from the supernatant formed in step (b) using a final alcohol concentration of from 24% to 26% at a pH of from 5.8 to 6.2.33. The method of claim 28 , wherein the method further comprises the step of:(c) precipitating Factor H from the supernatant formed in step (b) using a final alcohol concentration of from 20% to 30% at a pH of from 5.0 to 7.0.34. The method of claim 28 , wherein ...

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09-05-2013 дата публикации

PURIFICATION OF PROTEINS

Номер: US20130116413A1
Принадлежит:

The invention describes a method for protein purification. More particularly, the invention relates to a purification process comprising protein A chromatography and anion exchange chromatography wherein protein A chromatography eluate is further purified by anion exchange chromatography at similar pH or at a pH less than or equal to 6. 1. A process for purifying an antibody , comprising:a) purifying using protein A chromatography, wherein the antibody is eluted at low pH; andb) purifying using anion exchange chromatography performed in the flow-through mode, wherein eluate obtained from step a) is loaded onto an anion exchange resin at pH values less than or equal to 6.2. A process according to claim 1 ,wherein the antibody is eluted in step a) at pH values about 3.3 to about 4.5 and loaded onto the anion exchange resin at pH values about 3.3 to about 6.3. A process according to claim 1 , wherein the antibody is eluted in step a) at pH values about 3.5 and loaded onto the anion exchange resin at pH values about 3.5 to about 6.4. A process according to claim 1 , wherein the antibody is loaded onto the anion exchange resin at pH values about 4.5. A process according to claim 1 , wherein the antibody is loaded onto the anion exchange resin at pH values about 6.6. A process according to claim 1 , wherein the anion exchange chromatography is followed by a cation exchange chromatography step in a bind-elute mode claim 1 , and wherein the flow-through from the anion exchange chromatography step is loaded onto a cation exchange resin at pH values less than or equal to 6.7. A process for purifying an antibody claim 1 , comprising:a) purifying using protein A chromatography, wherein the antibody is eluted at low pH values; andb) purifying using anion exchange chromatography, performed in the flow-through mode, wherein eluate obtained from step a) is loaded on to the anion exchange resin without substantial adjustment of pH. Aspects of this application relate to purification ...

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16-05-2013 дата публикации

CYCLOSPORIN A FORM 2 AND METHOD OF MAKING SAME

Номер: US20130123193A1
Автор: Smith Scott W., WU Ke
Принадлежит: ALLERGAN, INC.

Disclosed herein are methods of making cyclosporin A Form 2. 1. A method for making cyclosporin A form 2 , the method comprising the steps of:a) preparing a suspension of cyclosporin A in a solvent comprising water and an ingredient selected from the group consisting of acetonitrile, 1,4-dioxane, and ethanol;b) a first heating-cooling cycle comprising heating the suspension to a temperature of between about 5° C. and about 50° C., followed by cooling the suspension to a temperature of about 1° C. to about 35° C.;c) a second heating-cooling cycle comprising heating the suspension to a temperature of between about 5° C. and about 50° C., followed by cooling the suspension to a temperature of about 1° C. to about 35° C.; andd) a third heating-cooling cycle comprising heating the suspension to a temperature of between about 5° C. and about 50° C., followed by cooling the suspension to a temperature of about 1° C. to about 35° C.2. The method of claim 1 , wherein the solvent comprises water and acetonitrile.3. The method of claim 2 , wherein the molar fraction of water in the solvent is between about 0.8 and about 1.0.4. The method of claim 3 , wherein the molar fraction of water in the solvent is about 0.87.5. The method of claim 1 , wherein the solvent comprises water and 1 claim 1 ,4-dioxane.6. The method of claim 5 , wherein the molar fraction of water in the solvent is between about 0.8 and about 1.0.7. The method of claim 6 , wherein the molar fraction of water in the solvent is about 0.90.8. The method of claim 1 , wherein the solvent comprises water and ethanol.9. The method of claim 8 , wherein the molar fraction of water in the solvent is between about 0.8 and about 1.0.10. The method of claim 9 , wherein the molar fraction of water in the solvent is about 0.89.11. The method of any of claim 1 , wherein the first claim 1 , second claim 1 , or third heating-cooling cycle comprises heating the suspension to a temperature of between about 5° C. and about 50° C. at ...

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16-05-2013 дата публикации

Sustained action formulation of cyclosporin form 2

Номер: US20130123195A1
Принадлежит: Allergan Inc

Disclosed herein are methods of treating diseases of the eye by administering to the subconjunctival space a formulation comprising cyclosporin A form 2 and a hydrogel.

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16-05-2013 дата публикации

METHOD FOR EXTRACTING UNDENATURED TYPE II COLLAGEN HAVING ACTIVE EPITOPE

Номер: US20130123468A1
Принадлежит:

A method for extracting water-soluble undenatured type II collagen having an active epitope comprises a first step for extracting animal-derived cartilage using an acidic solution at 50° C. or less, and a second step for adding pepsin to the acidic solution and performing extraction at 40° C. or less. This allows the epitope of undenatured type II collagen to be extracted in large quantities and with high efficiency without any loss of activity, and makes it possible to provide a food or beverage product obtained using the water-soluble undenatured type II collagen extracted by the extraction method. 111-. (canceled)12. An extraction method wherein water-soluble undenatured type II collagen is extracted and an active epitope contained in the water-soluble undenatured type II collagen is extracted , comprising the steps of: (1) temperature: 5 to 35° C.,', '(2) pH: 3.4 or less, and', '(3) extraction time period: 12 to 36 hours; and, 'extracting animal-derived cartilage with an acidic solution under the following conditions (1) temperature: 5 to 35° C.,', '(2) pH: 3.4 or less, and', '(3) extraction time period: 12 to 36 hours., 'adding pepsin to the acidic solution and extracting an active epitope with the acidic solution under the following conditions13. The extraction method according to claim 12 , wherein the step of extracting animal-derived cartilage with the acidic solution is a first step and the step of adding pepsin to the acidic solution and extracting an active epitope with the acidic solution is a second step claim 12 , and the total extraction time for both the first step and the second step is 48 hours or less.14. The extraction method according to claim 13 , wherein the second step is performed at lower temperature than the first step.15. The extraction method according to claim 13 , wherein elastase is further added in the second step of extraction.16. The extraction method according to claim 14 , wherein elastase is further added in the second step of ...

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16-05-2013 дата публикации

Purification of Proteins

Номер: US20130123476A1
Автор: Moya Wilson
Принадлежит: EMD MILLIPORE CORPORATION

The present invention relates to a selectively soluble polymer capable of binding to one or more constituents in a mixture containing various biological materials and the methods of using such a polymer to purify a biomolecule from such a mixture. The polymer is soluble in the mixture under a certain set of process conditions such as pH or temperature and is rendered insoluble and precipitates out of solution upon a change in the process conditions. While in its solubilized state, the polymer is capable of binding to a selected entity within the stream such as impurities (DNA, RNA, host cell protein, endotoxins, etc) in a cell broth and remains capable of binding to that entity even after the polymer is precipitated out of solution. The precipitate can then be filtered out from the remainder of the stream and the desired biomolecule is recovered and further processed. 1. The method of purifying a biomolecule of interest from a mixture containing the biomolecule of interest and one or more impurities , wherein the biomolecule of interest is produced in a microbial cell , the method comprising the steps of:a) providing the mixture containing a biomolecule of interest and one or more impurities, wherein the biomolecule is expressed in a microbial cell;b) adjusting the pH of the mixture;c) adding a polymer to the mixture, wherein the polymer is selected from PVP and a co-polymer of PVP;d) increasing the pH of the mixture, to precipitate the polymer and form a supernatant, wherein the precipitated polymer binds the one or more impurities; ande) recovering the supernatant containing the biomolecule of interest, thereby to purify the biomolecule of interest.2E.coli. The method of claim 1 , wherein the microbial cell is an cell.3. The method of claim 1 , wherein the microbial cell is a yeast cell.4. The method of claim 1 , wherein the pH is adjusted to a pH of about 4.5 in step (b).5. The method of claim 1 , wherein the pH is increased to a pH ranging from 5-7 in step (d).6 ...

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23-05-2013 дата публикации

Purification of anti-c-met antibodies

Номер: US20130129718A1
Принадлежит: Genentech Inc

Provided herein are methods of purifying anti-c-met antibodies, compositions and pharmaceutical formulations comprising purified anti-c-met antibodies, and methods of using the same.

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23-05-2013 дата публикации

PREPARATION OF CANOLA PROTEIN ISOLATE FROM CANOLA OIL SEEDS ("BLENDERTEIN")

Номер: US20130131316A1
Принадлежит:

Canola protein isolate is recovered from canola oil seeds by crushing the oil seeds and extracting the crushed canola oil seeds. Fat co-extracted from the crushed oil seeds is removed from the aqueous canola protein solution which then is processed by the micellar route to obtain the canola protein isolate. 1. A process for the preparation of a canola protein isolate , which comprises:grinding canola oil seeds,extracting the ground canola oil seeds with an aqueous extracting medium to solubilize canola protein and fats in the ground canola oil seeds to form an aqueous canola protein solution,separating the aqueous canola protein solution from residual ground canola oil seeds,defatting the aqueous canola protein solution,clarifying the defatted aqueous canola protein solution,concentrating the clarified aqueous canola protein solution while maintaining the ionic strength substantially constant to form a concentrated canola protein solution,diluting the concentrated protein solution into chilled water to cause the formation of canola protein micelles,collecting the canola protein micelles as a protein micellar mass, anddrying the protein micellar mass to form a canola protein isolate having a protein content of at least about 90 wt % (N×6.25) d.b.2. The process of claim 1 , wherein said aqueous extracting medium is an aqueous salt solution having an ionic strength of at least about 0.05 M with a pH of about 5 to about 6.8 to form a canola protein solution having a concentration of about 3 to about 40 g/L.3. The process of wherein an antioxidant is present in the aqueous extracting medium.4. The process of wherein said defatting step is effected by chilling the canola protein solution to a temperature of about 3° to about 7° C. and removing fat that separates from the canola protein solution.5. The process of wherein claim 4 , following the defatting step claim 4 , the separated aqueous canola protein solution is subjected to a colour removal step.6. The process of ...

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30-05-2013 дата публикации

Purification Of Proteins

Номер: US20130137860A1
Автор: Jaber Jad, Moya Wilson
Принадлежит: EMD MILLIPORE CORPORATION

The present invention relates to a selectively soluble polymer capable of binding to a desired biomolecules in a mixture containing various biological materials and the methods of using such a polymer to purify a biomolecule from such a mixture. The polymer is soluble in the mixture under a certain set of process conditions such as pH or temperature and/or salt concentration and is rendered insoluble and precipitates out of solution upon a change in the process conditions. The polymer is capable of binding to the desired biomolecule (protein, polypeptide, etc) and remains capable of binding to that biomolecule even after the polymer is precipitated out of solution. The precipitate can then be filtered out from the remainder of the stream and the desired biomolecule is recovered such as by elution and further processed. 1. A method for purifying a recombinant protein produced in a microbial host cell , the method comprising the steps of:a. providing a mixture comprising a recombinant protein produced in a microbial host cell;b. adding one or more stimuli responsive polymers to the mixture under pH conditions such that the one or more polymers are solubilized and are capable of reversibly and selectively binding to the recombinant protein;c. mixing the one or more solubilized stimuli responsive polymers throughout the mixture;d. changing pH conditions such that to precipitate the one or more polymers and bound recombinant protein out of solution; and e. separating the precipitated polymer and bound protein from the mixture.2. The method of claim 1 , wherein the one or more stimuli responsive polymers are one or more cationic polyelectrolytes.3. The method of claim 2 , wherein the one or more cationic polyelectrolytes is selected from the group consisting of chitosan claim 2 , polyvinylpyridine claim 2 , copolymers of vinylpyridine claim 2 , primary amine containing polymers claim 2 , secondary amine containing polymers and tertiary amine containing polymers.4. The ...

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06-06-2013 дата публикации

PROTEIN FRACTIONATION BASED ON pI

Номер: US20130140182A1

Methods and devices for detecting and collection analytes fractionated based on pI, separating analytes via electrophoresis and pI, and purifying a target molecule using pI focusing and subsequent crystallization are provided.

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13-06-2013 дата публикации

Crystalline Peptide Epoxy Ketone Protease Inhibitors and the Synthesis of Amino Acid Keto-Epoxides

Номер: US20130150289A1
Принадлежит: Onyx Therapeutics, Inc.

The invention relates to crystalline peptide keto epoxide compounds, methods of their preparation, and related pharmaceutical compositions. This invention also relates to methods for the preparation of amino acid keto-epoxides. Specifically, allylic ketones are stereoselectively converted to the desired keto epoxides. 118.-. (canceled)20. A method of claim 19 , wherein the organic solvent is selected from diethyl ether claim 19 , THF claim 19 , acetonitrile claim 19 , and MTBE claim 19 , or any combination thereof.21. A method of claim 20 , wherein the organic solvent is a mixture of THF and acetonitrile.22. A method of claim 19 , wherein bringing the solution to supersaturation comprises slow addition of an anti-solvent claim 19 , allowing the solution to cool claim 19 , reducing the volume of the solution claim 19 , or any combination thereof.23. A method of claim 22 , wherein bringing the solution to supersaturation comprises cooling the solution to ambient temperature or lower.24. A method of claim 19 , further comprising washing the crystals.25. A method of claim 24 , wherein the washing comprises washing with a liquid selected from diethyl ether claim 24 , THF claim 24 , acetonitrile claim 24 , and MTBE claim 24 , or any combination thereof.26. A method of claim 25 , wherein washing comprises washing with acetonitrile.27. A method of claim 19 , wherein isolating the crystals comprises filtering the crystals.28. A method of claim 19 , further comprising drying the crystals under reduced pressure.30. A crystalline salt of claim 29 , having a DSC thermogram substantially as shown in .31. A crystalline salt of claim 29 , having a melting point of about 180 to about 190° C.32. A crystalline salt of claim 31 , having a melting point of about 184 to about 188° C.33. A crystalline salt of claim 29 , having an XRPD pattern substantially as shown in .34. A crystalline salt of claim 29 , having 2θ values 4.40; 7.22; 9.12; 12.36; 13.35; 14.34; 15.54; 16.14; 16.54; 17.00; ...

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27-06-2013 дата публикации

PROTEIN SEPARATION VIA ION-EXCHANGE CHROMATOGRAPHY AND ASSOCIATED METHODS, SYSTEMS, AND DEVICES

Номер: US20130164827A1
Автор: Torres Anthony R.
Принадлежит:

The present invention provides methods for separating proteins from a protein mixture. In one aspect, a method for separating a high concentration protein mixture into a bound protein fraction and a flow-through protein fraction can include delivering a protein mixture through an ion exchange column at a fixed pH and a fixed salt concentration. The fixed pH and the fixed salt concentration have been preselected to cause separation of the protein mixture into a bound protein fraction and a flow-through protein fraction. In this case, the bound protein fraction binds to the ion exchange column and the flow-though protein fraction flows though the ion exchange column. The method can further include receiving the flow-through protein fraction from the ion exchange column separate from the bound protein fraction, wherein either the bound protein fraction or the flow-through fraction contains a protein of interest. 1. A method for separating a high concentration protein mixture into a bound protein fraction and a flow-through protein fraction , comprising:delivering a high concentration protein mixture through an ion exchange column at a fixed pH and a fixed salt concentration, wherein the fixed pH and the fixed salt concentration have been preselected to cause separation of the high concentration protein mixture into a bound protein fraction and a flow-through protein fraction, wherein the bound protein fraction binds to the ion exchange column and the flow-though protein fraction flows though the ion exchange column; andreceiving the flow-through protein fraction from the ion exchange column separate from the bound protein fraction, and wherein either the bound protein fraction or the flow-through fraction contains a protein of interest.2. The method of claim 1 , wherein the protein of interest is in the flow-through protein fraction.3. The method of claim 1 , wherein the protein of interest is in the bound protein fraction.4. The method of claim 1 , wherein the high ...

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04-07-2013 дата публикации

Intravenous Cytomegalovirus Human Immune Globulin and Manufacturing Method Thereof

Номер: US20130172536A1

The present invention discloses an intravenous cytomegalovirus human immune globulin and a manufacturing method thereof, wherein the technical problem to be solved is to improve the purity, yield, and safety of the product. The intravenous cytomegalovirus human immune globulin of the present invention has a specific activity of no less than 2.5 PEI-U/mg, an anti-CMV titer of no less than 100 PEI-U/ml, a purity of greater than 98.2%, and a protein content of 51˜55 mg/ml. The present invention employs caprylic acid precipitation and anion exchange chromatography for replacing the step of ethanol precipitation in the conventional cold ethanol method to keep IgG in the supernatant and maintain the activity of the IgG; the present invention employing processes of caprylic acid inactivation of virus and nanometer film virus removal can effectively protect the safety of the product, and studies show that the preparing method of the present invention not only improves the purity, yield, and safety of the product; but also saves energy and reduces the cost of production. 1. An intravenous cytomegalovirus human immune globulin having a specific activity of no less than 2.5 PEI-U/mg , an anti-CMV titer of no less than 100 PEI-U/ml , a purity of greater than 98.2% , and a protein content of 51˜55 mg/ml.2. A method for preparing intravenous cytomegalovirus human immune globulin , comprising the steps of:(a) preparing FI+II+III and FII+III deposits:preparing human plasma measured by enzyme linked immunosorbent assay, dissolving said human plasma at 2˜30° C., and mixing said human plasma, wherein said human plasma has a high titer of anti-CMV;preparing FI+II+III deposit;adjusting the protein content of said human plasma to 45˜55 mg/ml with saline, adjusting the pH of said human plasma to 6.0˜6.5 with glacial acetic acid, and adding 95% ethanol or ethanol to adjust the concentration of ethanol of said human plasma to 20˜25%, wherein the reaction temperature is −5.5˜−4.5° C., and ...

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11-07-2013 дата публикации

PROTEIN PURIFICATION BY ION EXCHANGE

Номер: US20130178608A1
Принадлежит:

The application describes an antibody purification method comprising multiple chromatographic steps wherein the low pH eluate from a protein A chromatography is further purified without the need of substantial pH adjustment. 1. A process for purifying antibodies comprising:a) purifying using protein A chromatography, wherein the antibody is eluted at a particular pH value; andb) purifying using cation exchange chromatography in the bind-elute mode, wherein an eluate from step a) is loaded onto a cation exchange resin without substantial adjustment of pH.2. A process according to claim 1 , wherein the antibody is eluted in step a) at pH values about 3.3 to about 4.5.3. A process according to claim 1 , wherein the antibody is eluted in step a) at a pH value about 3.5.4. A process for purifying antibodies claim 1 , comprising:a) purifying using protein A chromatography, wherein an antibody is eluted at a first pH value;b) purifying using cation exchange chromatography performed in the bind-elute mode, wherein eluate obtained from step a) is loaded onto a cation exchange resin at a second pH value, and elution is carried out at a third pH value; andc) purifying using anion exchange chromatography performed in the flow-through mode, wherein eluate from step b) is loaded on to an anion exchange resin without substantial adjustment of pH.5. A process according to claim 4 , wherein the first and second pH values are similar.6. A process according to claim 4 , wherein the first and second values are different.7. A process according to claim 4 , wherein the second and third pH values are similar.8. A process according to claim 4 , wherein the first pH value is about 3.59. A process according to claim 4 , wherein the second pH value is about 6 to about 8.10. A process according to claim 4 , wherein the second pH value is about 6.11. A process according to claim 4 , wherein the third pH value is about 6 to about 8.12. A process according to claim 4 , wherein the third pH value ...

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18-07-2013 дата публикации

METHOD FOR ISOLATING A CYCLOHEXAPEPTIDE

Номер: US20130184433A1
Принадлежит: DSM Sinochem Pharmaceuticals

The present invention relates to a method for isolating acyclohexapeptide and to a novel crystalline form of caspofungin diacetate thus obtained. 1. Method for isolating a cyclohexapeptide comprising the subsequent steps of:(a) Contacting a solution comprising a cyclohexapeptide and water with a resin;(b) Removing the liquid phase from the mixture obtained in step (a);(c) Eluting said cyclohexapeptide by contacting the material retained after removal of the liquid phase in step (b) by addition of an organic solvent;(d) Separating resin from the mixture obtained in step (c);(e) Crystallizing said cyclohexapeptide from the solution retained after removal of said resin(f) Isolating said cyclohexapeptide from the mixture obtained in step (e)2. Method according to wherein said resin is contained in a chromatography column claim 1 , wherein step (a) is carried out by introducing said solution comprising a cyclohexapeptide and water onto said chromatography column and wherein step (b) is carried out by removing said liquid phase from said chromatography column.3. Method according to wherein said resin is washed with an aqueous mixture after step (a) and before step (c).4. Method according to wherein said resin is hydrophobic interaction chromatography material claim 1 , reversed phase chromatography material claim 1 , ion-exchange chromatography material or a mixture of two or more of these materials.5. Method according to wherein said organic solvent in step (c) is an alcohol.6. Method according to wherein said cyclohexapeptide is anidulafungin or a salt thereof or caspofungin or a salt thereof or micafungin or a salt thereof.7. Method according to wherein the concentration of said cyclohexapeptide in said solution of step (a) is from 2 to 50 times lower than the concentration of said cyclohexapeptide in the solution retained after removal of said resin in step (d).8. Method according to wherein the concentration of said cyclohexapeptide in said solution of step (a) is ...

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18-07-2013 дата публикации

METHOD FOR OBSERVING PROTEIN CRYSTAL

Номер: US20130184445A1
Принадлежит: OSAKA UNIVERSITY

Provided are a method for observing protein crystal, wherein the growth process of the protein crystals is nondestructively and three-dimensionally monitored on a real-time basis and the growth of the crystals is controlled at a high accuracy to thereby enable the formation of single crystals having good qualities, which comprises observing the protein crystals, said protein crystals having been produced by a crystallization method using a gel, by an OCT measurement using light emitted from an ultrawideband light source; 1. A method for observing protein crystal for producing a good quality crystal of a protein , wherein protein crystal produced by a crystallization method using a gel is observed by OCT measurement measuring light emitting from an ultra-wideband light source and an interfering light generated from a scattering light of the emitting light.2. The method for observing protein crystal according to claim 1 , wherein the above-described ultra-wideband light source is an ultra-wideband supercontinuum light source.3. The method for observing protein crystal according to claim 2 , wherein the center wavelength of light emitting from the above-described ultra-wideband supercontinuum light source is of 0.8 μm band.4. The method for observing protein crystal according to claim 1 , wherein the above-described observation for protein crystal is observation by in-situ measurement.5. A method for evaluating protein crystal claim 1 , wherein evaluation of protein crystal is carried out using the method for observing protein crystal according to .6. A method for distinguishing a protein crystal and a salt claim 1 , wherein separation of a protein crystal and a salt is carried out using the method for observing protein crystal according to .7. A method for separating a protein crystal and a salt for producing quality crystal of a comprisinga gel liquid production step of gelling a material to be measured containing a protein and a salt to produce gel liquid,a protein ...

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25-07-2013 дата публикации

COLLAGEN POWDER AND/OR COLLAGEN-DERIVED POWDER, AND PRODUCTION METHOD FOR THE SAME

Номер: US20130190479A1
Принадлежит: NIPPI, INCORPORATED

Disclosed is a collagen powder and/or a collagen derivative powder, which are obtained by dispersing in a hydrophilic organic solvent a crude collagen precipitate which comprises 12 to 50% by mass of a collagen precipitate and/or a collagen derivative precipitate having an average particle size of 1 to 1,000 μm, recovering solids and then drying the solids. By dispersing the crude collagen precipitate in the hydrophilic organic solvent, the resulting precipitates can be dehydrated, so that drying of the thus obtained solids can be done by air-drying. In addition, the resulting collagen powder and/or collagen derivative powder exhibit excellent solubility due to an increased specific surface area and also have excellent ease of handling with the average particle size being 8 to 1,000 μm. 1. A collagen powder and/or a collagen derivative powder , which are obtained by dispersing in a hydrophilic organic solvent a crude collagen precipitate which comprises 12 to 50% by mass of a collagen precipitate and/or a collagen derivative precipitate having an average particle size of 1 to 1 ,000 μm , recovering solids and then drying the solids.2. The collagen powder and/or the collagen derivative powder according to claim 1 , which is characterized in that the crude collagen precipitate is an isoelectric precipitate of a collagen and/or a collagen derivative obtained at a pH of 3.5 to 10 or a salt precipitate.3. The collagen powder and/or the collagen derivative powder according to claim 1 , which is characterized in that:the crude collagen precipitate has an isoelectric point at a pH of 3.5 to 8.0; andthe initial dissolution rate of a pH 6.5 solution comprising 5 mg of the powder is not less than 0.2 mg/min.4. A method of producing a collagen powder and/or a collagen derivative powder claim 1 , which is characterized by comprising the steps of:obtaining isoelectric precipitates having an average particle size of 1 to 1,000 μm by isoelectrically precipitating a solution ...

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01-08-2013 дата публикации

ULTRAFILTRATION AND DIAFILTRATION FORMULATION METHODS FOR PROTEIN PROCESSING

Номер: US20130195888A1
Принадлежит: AbbVie

Disclosed herein are methods of purifying proteins using ultrafiltration and diafiltration processes. 1. A method for preparing a formulation comprising the steps of:a) providing a first solution, said first solution comprising one or more proteins;b) subjecting said first solution to diafiltration, using a diafiltration solution, said diafiltration solution comprising water, until at least a five-fold volume exchange with said diafiltration solution has been achieved, thereby forming a second solution; andc) concentrating said one or more proteins in said second solution within a range of about 10 grams per liter to about 300 grams per liter.2. The method of claim 1 , further comprising:d) adding one or more buffer salts and/or one or more excipients to the solution of step c).3. The method of claim 1 , wherein said diafiltration solution further comprises a surfactant.4. The method of claim 3 , wherein said diafiltration solution comprises about 0.0001 percent to about 0.5 percent (w/v) polysorbate.5. The method of claim 1 , further comprising adjusting the pH of said first solution within a range of about 4 to about 6.6. The method of claim 5 , wherein said first solution further comprises acetic acid and/or citric acid in an amount sufficient to adjust the pH of said first solution within a range of about 4 to about 6.7. The method of claim 1 , wherein said concentrating step c) comprises ultrafiltering said second solution.8. The method of claim 2 , wherein said one or more buffer salts comprises histidine within a concentration range of about 10 millimolar to about 100 millimolar.9. The method of claim 2 , wherein histidine is added during any of steps a) through d) in a sufficient amount to achieve a target concentration of about 15 mM histidine at a target pH of about 6 in the formulation.10. The method of claim 1 , wherein said one or more proteins is an antibody claim 1 , or antigen binding fragment thereof.11. The method of claim 10 , wherein the antibody ...

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01-08-2013 дата публикации

METHOD FOR FORMING PROTEIN CRYSTAL

Номер: US20130196160A1
Автор: Sugahara Michihiro
Принадлежит: RIKEN

A method for producing a capsule for protein crystallization is provided. The method comprises adding a solution containing a protein and a gelling agent to an ionic cross-linking solution to form an ionically cross-linked gel capsule that encapsulates a solution of the protein. 1. A method for producing a capsule for protein crystallization , comprising:adding a solution containing a protein and a gelling agent to an ionic cross-linking solution to form an ionically cross-linked gel capsule that encapsulates a solution of the protein.2. A composition for protein crystallization comprising a shell including an ionically cross-linked gel , wherein the shell encapsulates a solution including a protein.3. A kit for producing a capsule for protein crystallization , comprising a gelling agent and an ionic cross-linking solution.4. The kit for producing a capsule for protein crystallization according to claim 3 , further comprising:a crystallization solution;a non-volatile oil; anda plate for protein crystallization, wherein the plate has one or more wells for placing the crystallization solution and the non-volatile oil therein.5. The kit for producing a capsule for protein crystallization according to claim 4 , wherein the cross-linking solution and the crystallization solution are identical to each other.6. The kit for producing a capsule for protein crystallization according to claim 4 , wherein the crystallization solution and the non-volatile oil are placed in each well of the plate claim 4 , and the kit further comprises a film for sealing the well.7. A method for producing a protein crystal claim 4 , comprising:(a) adding a solution containing a protein and a gelling agent to an ionic cross-linking solution to form an ionically cross-linked gel capsule; and(b) maintaining the capsule formed in step (a) in a crystallization solution.8. The method for producing a protein crystal according to claim 7 , wherein the ionic cross-linking solution and the crystallization ...

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01-08-2013 дата публикации

METHOD FOR PRODUCING SURFACTIN AND SALT THEREOF

Номер: US20130197190A1
Принадлежит: KANEKA CORPORATION

The method for producing a surfactin or a salt thereof according to the present invention comprises the steps of adding an organic solvent containing a branched alkyl alcohol to a culture fluid containing the surfactin or the salt thereof, or to a solution obtained by removing an insoluble component from the culture fluid, and extracting the surfactin or the salt thereof with the organic solvent, 2. The method according to claim 1 , wherein an aqueous layer during the extraction is acidic.3. The method according to claim 2 , wherein a pH value of the aqueous layer during the extraction is not less than 1 and not more than 5.4. The method according to claim 1 , wherein a carbon number of the branched alkyl alcohol is not less than 3 and not more than 10.5. The method according to claim 1 , wherein a ratio of the branched alkyl alcohol in the organic solvent is not less than 30 wt %.6. The method according to claim 1 , wherein the organic solvent further contains an auxiliary solvent claim 1 , and a total ratio of the branched alkyl alcohol and the auxiliary solvent is not less than 0.6 parts by weight and not more than 1.5 parts by weight relative to 1 part by weight of the culture fluid.7. The method according to claim 1 , further comprising the steps of:a) obtaining an aqueous solution containing the surfactin salt by mixing the extract containing the surfactin with a basic aqueous solution and then removing an organic layer; andb) mixing the aqueous solution containing the surfactin salt with an inorganic acid to precipitate the surfactin in a solid form from the obtained mixture.8. The method according to claim 1 , further comprising the step of mixing the extract containing the surfactin with a base to obtain the surfactin salt obtained in a solid form from the obtained mixture.9. A method for producing the surfactin salt claim 1 , comprising the step of obtaining the surfactin salt or a solution thereof by mixing the surfactin obtained by the method according ...

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01-08-2013 дата публикации

METHODS OF REDUCING LEVEL OF ONE OF MORE IMPURITIES IN A SAMPLE DURING PROTEIN PURIFICATION

Номер: US20130197200A1
Принадлежит: EMD MILLIPORE CORPORATION

The present invention provides novel and improved protein purification processes which incorporate certain types of carbonaceous materials and result in effective and selective removal of certain undesirable impurities without adversely effecting the yield of the desired protein product. 1. A flow-through process for purifying a target molecule from a Protein A eluate comprising the steps of:(i) contacting the eluate recovered from a Protein A chromatography column with activated carbon;(ii) contacting the flow-through sample from step (i) with an anion exchange chromatography media;(iii) contacting the flow-through sample from step (ii) with a cation exchange chromatography media; and(iv) obtaining the flow-through sample from step (iii) comprising the target molecule,wherein the eluate flows continuously through steps (i)-(iii) and wherein level of one or more impurities in the flow-through sample after step (iii) is lower than the level in the eluate in step (i).2. The flow-through process of claim 1 , further comprising subjecting the flow-through sample from step (iii) to virus filtration.3. The flow-through process of claim 1 , further comprising use of an in-line static mixer and/or a surge tank between steps (ii) and (iii) to change pH.4. The flow-through process of claim 1 , wherein the process employs a single skid.5. The flow-through process of claim 1 , wherein the eluate from the Protein A chromatography column is subjected to virus inactivation prior to contacting with activated carbon.6. The process of claim 1 , wherein steps (i)-(iii) may be performed in any order.7. A flow-through purification process for purifying a target molecule from a Protein A eluate comprising contacting the eluate with two or more matrices selected from activated carbon claim 1 , anion exchange media claim 1 , cation exchange media and virus filtration media claim 1 , wherein the flow of the eluate is continuous.8. A method for reducing the burden on one or more ...

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08-08-2013 дата публикации

Flexible dry sprinklers

Номер: US20130199803A1
Автор: Thomas Multer
Принадлежит: Reliable Automatic Sprinkler Co Inc

A dry sprinkler is provided having a flexible tube section having openings at opposite ends of the flexible tube section. The sprinkler includes a supply line connection at one end of the tube section, the supply line connection having an opening to receive fluid from the supply line. The sprinkler has a first seal sealed with the supply line connection to prevent fluid from the supply from entering the tube section. A sprinkler head is coupled to the tube section at another end of the tube section opposite the supply line connection, the sprinkler head having a deflector, a second seal sealing an output orifice of the sprinkler head, and a thermally responsive element arranged to maintain the second seal sealed. A fluid is contained in the tube between the first seal and the second seal, wherein the fluid is not susceptible to freezing. The thermally responsive element is constructed to release the second seal in response to an elevated temperature condition, to permit the release of the fluid in the tube and release the first seal to move toward the sprinkler head.

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08-08-2013 дата публикации

METHOD FOR PURIFICATION OF COMPLEMENT FACTOR H

Номер: US20130203971A1
Принадлежит: Octapharma AG

A method for purification of complement Factor H from a complement Factor H containing source such as blood or blood plasma, in particular a caprylate precipitate of a Factor H containing source, which is e.g. obtained by addition of caprylate ions to fractions of blood or plasma, comprising the steps of: 1. A method for purification of complement Factor H from a complement Factor H containing source , comprising the steps of:a. Providing a Factor containing source;b. performing a cation exchange chromatography;c. performing an anion exchange chromatography;d. performing a hydroxyl apatite chromatography;e. followed by ultra/diafiltration to obtain a complement Factor H concentrate.2. The method according to wherein a heparin affinity chromatography is performed.3. The method according to wherein the process comprises at least one of the following methods for pathogen removal and/or inactivation:a. solvent/detergent treatment;b. pasteurization,c. vapor heat treatment;d. dry heat treatment; ore. nanofiltration.4. The method according to wherein the complement Factor H concentrate is lyophilized.5. A complement Factor H obtainable by a method according to .6. The complement Factor H of obtainable by a method comprisinga. reconstitution of caprylate precipitate to provide a complement Factor H containing solution;b. performing virus inactivation by solvent/detergent treatment (S/D treatment).c. performing a cation exchange chromatography chromatographic step under the following conditions:d. binding of complement Factor H to a strong cation exchange resin of the sulphopropyl type, washing with a buffer comprising 20 mM tri-sodium citrate adjusted to pH 6.0 and elution of complement Factor H with an elution buffe r comprising 20 mM tri-sodium citrate and 0.2 M NaCl adjusted to pH 6.0; 'i. applying of the complement Factor H containing solution (conductivity 0.1-0.5 mS/cm) to a strong anion exchange resin of the quaternary ammonium type, washing with a buffer comprising ...

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15-08-2013 дата публикации

METHOD FOR PURIFYING HUMAN GRANULOCYTE-COLONY STIMULATING FACTOR FROM RECOMBINANT E. COLI

Номер: US20130211054A1
Принадлежит: HANMI SCIENCE CO., LTD.

The present invention provides a method for purifying a large amount of human granulocyte-colony stimulating factors (hG-CSFs) from a recombinant with high yield and purity. According to the method of the present invention, human granulocyte-colony stimulating factor, identical to the native form expressed in the human body, can be easily purified with high yield and purity without an additional activation process. In particular, according to the purification method of the present invention, hG-CSF variants expressed in are efficiently removed to obtain physiologically active hG-CSFs with high purity. 1E. coli. A method for purifying human granulocyte-colony stimulating factors (hG-CSFs) from a recombinant in a high yield , comprising the steps of:{'i': 'E. coli', '(a) culturing an hG-CSF-expressing recombinant to obtain a cell pellet by centrifugation;'}(b) separating an hG-CSF-containing supernatant from the cell pellet obtained in step (a);(c) treating the supernatant obtained in step (b) with an acid to separate the resulting precipitate by filtration;(d) applying a filtrate obtained in step (c) to cation exchange chromatography;(e) applying an eluate obtained in step (d) to hydrophobic-interaction chromatography; and(f) applying an eluate obtained in step (e) to anion exchange chromatography.2E. coli.. The method according to claim 1 , wherein in step (a) claim 1 , the hG-CSFs are expressed into the periplasm of recombinant3E. coliE. coliE. coliE. coliE. coliE. coliE. coliE. coliE. coliE. coliE. coliE. coli. The method according to claim 2 , wherein the recombinant is one or more selected from the group consisting of BL21(DE3)/pT14SS1SG(HM 10310) claim 2 , BL21(DE3)/pT14SS1S17SEG (HM 10311; Accession No. KCCM-10154) claim 2 , BL21(DE3)/pTO1SG (HM 10409) claim 2 , BL21(DE3)/pTO1S17SG (HM 10410; Accession No. KCCM-10151) claim 2 , BL21(DE3)/pTO17SG (HM 10411; Accession No. KCCM-10152) claim 2 , BL21(DE3)/pTO17TG (HM 10413) claim 2 , BL21(DE3)/pTO17AG(HM 10414) ...

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29-08-2013 дата публикации

METHOD TO PRODUCE AN IMMUNOGLOBULIN PREPARATION WITH IMPROVED YIELD

Номер: US20130224184A1
Принадлежит:

The present invention provides improved methods for the manufacturing of IVIG products. These methods offer various advantages such as reduced loss of IgG during purification and improved quality of final products. In other aspects, the present invention provides aqueous and pharmaceutical compositions suitable for intravenous, subcutaneous, and/or intramuscular administration. In yet other embodiments, the present invention provides methods of treating a disease or condition comprising administration of an IgG composition provided herein. 1. A method for preparing an enriched IgG composition from plasma , the method comprising the steps of:(a) precipitating a cryo-poor plasma fraction, in a first precipitation step, with between about 6% and about 10% alcohol at a pH of between about 7.0 and about 7.5 to obtain a first precipitate and a first supernatant;(b) precipitating IgG from the first supernatant, in a second precipitation step, with between about 20% and about 25% alcohol at a pH of between about 6.7 and about 7.3 to form a second precipitate;(c) re-suspending the second precipitate to form a suspension;(d) precipitating IgG from the suspension formed in step (c), in a third precipitation step, with between about 22% and about 28% alcohol at a pH of between about 6.7 and about 7.3 to form a third precipitate;(e) re-suspending the third precipitate to form a suspension; and(f) separating the soluble fraction from the suspension formed in step (e), thereby forming an enriched IgG composition,wherein the pH of at least one of the first precipitation step, second precipitation step, or third precipitation step is achieved by addition of a pH modifying agent after addition of the alcohol.2. (canceled)3. (canceled)4. The method of claim 1 , wherein the pH of all of the first precipitation step claim 1 , second precipitation step claim 1 , and third precipitation step is achieved by addition of a pH modifying solution after addition of the alcohol.5. The method of ...

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05-09-2013 дата публикации

Human placental collagen compositions, and methods of making and using the same

Номер: US20130231288A1
Принадлежит: Anthrogenesis Corp

The present invention provides compositions comprising human placental telopeptide collagen, methods of preparing the compositions, methods of their use and kits comprising the compositions. The compositions, kits and methods are useful, for example, for augmenting or replacing tissue of a mammal.

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03-10-2013 дата публикации

Continuous processing methods for biological products

Номер: US20130260419A1
Принадлежит: TARPON BIOSYSTEMS Inc

The present invention is directed to the development of continuous processing technology for the purification of biopharmaceuticals and biological products, such as monoclonal antibodies, protein therapeutics, and vaccines. Methods for continuous processing of a biological product in a feed stream toward formulation of a purified bulk product are described.

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03-10-2013 дата публикации

Method of co-extracting multiple proteins from chicken egg white

Номер: US20130260437A1
Автор: Geng Fang, HUANG QUN, Ma Meihu
Принадлежит: Huazhong Agriculture University

A method of co-extracting multiple proteins from chicken egg white is provided. Precipitate proteins step-by-step with polyethylene glycol, then separate the proteins via Q Sepharose Fast Flow anion-exchange chromatography. The method of co-extracting multiple proteins from chicken egg white is capable of obtaining five proteins simultaneously in one extraction process, and the protein products have high purity and high recovery ratio. And the method can be carried out only requiring common-used chromatographic filler, greatly reduces production cost, and provides favorable factors for large-scale industrial production of extracting protein from egg white. 1. A method of co-extracting multiple proteins from chicken egg white , comprising steps of:(1) diluting fresh egg white with 20˜100 mmol/L NaCl solution of an equal volume, mixing, and adjusting PH to 5.0˜8.0, so as to obtain processed egg white diluted solution;(2) adding polyethylene glycol-8000 (PEG-8000) to the egg white diluted solution while stirring, until mass percentage concentration of the PEG-8000 is 2˜6%, fully stirring for 2˜12 hours, and then processing centrifugation to obtain precipitate A and supernatant A,continually adding PEG-8000 to the supernatant A until mass percentage concentration of the PEG-8000 is 8˜12%, fully stirring for 2˜12 hours, then processing centrifugation to obtain precipitate B and supernatant B, andcontinually adding PEG-8000 to the supernatant B until mass percentage concentration of the PEG-8000 is 14˜18%, fully stirring for 2˜12 hours, then processing centrifugation to obtain precipitate C and supernatant D;(3) suspending the precipitate A in 200˜800 mmol/L NaCl solution, fully stirring, centrifuging after suspending for 4˜24 hours to obtain new precipitate, suspending the new precipitate in distilled water, fully stirring, centrifuging, suspending in distilled water again and processing centrifugation again, so as to wash away sodium chloride, collecting a precipitate, ...

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03-10-2013 дата публикации

METHOD FOR PREPARATIVE IN VITRO PROTEIN BIOSYNTHESIS

Номер: US20130261290A1
Принадлежит:

The invention relates to a method for preparative in vitro protein synthesis in a cell-free transcription/translation system, comprising the following steps: a) in a reaction vessel, a reaction solution is prepared, comprising the following synthesis substances: components of the transcription/translation apparatus for a defined-protein, amino acids, and metabolic components supplying energy and being necessary for the synthesis of the defined protein, b) the synthesis is performed in the reaction vessel in a defined period of time, c) after expiration of the defined period of time, the reaction solution is subjected to a separation step, in which generated low-molecular metabolic products are separated from the solution (and extracted). 1. A method for preparative in vitro protein synthesis of an expression product in a cell-free transcription/translation system , comprising the following steps:a) in a reaction vessel, a reaction solution is prepared, comprising the following synthesis substances: components of the transcription/translation apparatus for a defined expression product, amino acids, and metabolic components supplying energy and being necessary for the synthesis of the defined protein,b) the synthesis is performed in the reaction vessel in a defined period of time, without separating generated substances and without adding consumed synthesis substances within the defined period of time,c) after expiration of the defined period of time, the reaction solution is subjected to a separation step, in which generated low-molecular weight metabolic products and/or reaction inhibitors are separated from the solution,d) immediately before, after or at the same time as step c), consumed synthesis substances are supplemented,e) steps b), c) and d) are repeated at least once with the reaction solution of step d), wherein at the last execution of step b), the steps c) and d) may be omitted.2. The method according to claim 1 , wherein in step c) and/or subsequently ...

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17-10-2013 дата публикации

Methods for processing inclusion bodies

Номер: US20130273607A1
Автор: Ellen T. O'Connor
Принадлежит: MEDIMMUNE LLC

The present application relates to methods for purifying recombinant proteins, including antibodies and antibody fragments. Suitably, the methods utilize depth filtration to clarify the desired proteins from a solubilized mixture, and provide refolding methods and refolding buffers to allow for refolding of the recombinant proteins into functional and active proteins. Exemplary antibody fragments include anti-CD22 antibody fragments that comprise V H and V L chains refolded into a functional and active fragment.

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17-10-2013 дата публикации

PROCESS FOR PRODUCTION OF FIBRINOGEN

Номер: US20130274444A1
Принадлежит: Octapharma AG

The present invention relates to a method or process for the manufacture of a virus and prion save native fibrinogen concentrate of high purity and low amounts of fibrinopeptide A and fibronectin. 1. A process for purifying fibrinogen from a fibrinogen containing source , comprising subjecting a fibrinogen containing solution to chromatography on anion exchange resins , wherein the anion exchange resins are selected from the group consisting of a support material comprising a hydroxylated polymer with grafted tertiary or quaternary amino groups.2. The process of claim 1 , wherein the support material is composed of cellulose claim 1 , agarose claim 1 , silica claim 1 , polymeric or ceramic material.3. The process of claim 1 , wherein the anion-exchange resin is trimethyl-amino groups grafted to a hydroxylated methacrylic polymer via a linking group claim 1 , such as GigaCap Q-650M®.4. The process of claim 1 , wherein the fibrinogen containing source is cryoprecipitate claim 1 , preferably solubilised at about neutral pH .5. The process of claim 4 , wherein the solution is treated with Al(OH)and the resulting gel is removed.6. The process of wherein a virus inactivation is performed by employing a solvent/detergent (S/D) treatment.7. The process of wherein an extraction of S/D reagents with vegetable oil and contacting the water-phase with a TMAE resin at a pH-value of 6.9-7.1 and an osmolality of 570-610 mosmol/l is performed.8. The process of claim 6 , wherein fibrinogen is precipitated by glycine claim 6 , in particular about 1 M glycine claim 6 , and separation of the formed fibrinogen paste.9. The process of wherein the fibrinogen paste is resuspended claim 8 , preferably in about 20 mM TRIS buffer at a pH of about 8.0.10. The process of wherein after filtration the obtained fraction is loaded onto a strong anion exchange resin comprising trimethyl-amino groups grafted to a hydroxylated methacrylic polymer backbone via linking groups and washing off loosely ...

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24-10-2013 дата публикации

Purified antibody composition

Номер: US20130280267A1
Принадлежит: AbbVie Inc

The invention provides a method for producing a host cell protein-(HCP) reduced antibody preparation from a mixture comprising an antibody and at least one HCP, comprising an ion exchange separation step wherein the mixture is subjected to a first ion exchange material, such that the HCP-reduced antibody preparation is obtained.

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31-10-2013 дата публикации

METHODS FOR PURIFICATION OF ALPHA-1-ANTITRYPSIN AND APOLIPOPROTEIN A-1

Номер: US20130289245A1
Принадлежит: CSL Behring GmbH

This invention relates to protein separation and purification methods for both alpha-1-antitrypsin (AAT, also known as alpha-1 proteinase inhibitor, API, and A.sub.1-PI) and Apolipoprotein A-I (ApoA-1) from, for example, a fraction of human blood plasma. In certain embodiments, the invention provides methods for separating AAT from ApoA-1 at the initial stage of purification, so that the same starting material can be used as a source for both proteins. The methods further pertain to providing compositions of AAT and of ApoA-1 suitable for pharmaceutical use and are suitable for large-scale purification. 125.-. (canceled)26. A method for purifying Apolipoprotein A-I (ApoA-I) and Alpha-1-Antitrypsin (AAT) from a single starting human plasma fraction containing both proteins comprising: (a) treating the starting human plasma fraction which is used as the starting material such that ApoA-I and AAT are solubilized;', '(b) adding an ApoA-I binding agent and ensuring an appropriate pH so that ApoA-I binds to the added ApoA-I binding agent, wherein the binding agent is fumed silica;', '(c) separating ApoA-I bound to said fumed silica from the solution containing AAT;', '(d) eluting ApoA-I from said fumed silica; and, '(i) treating a starting human plasma fraction comprising ApoA-I and AAT to separate an ApoA-I containing fraction from an AAT containing fraction, comprising'}(ii) separately purifying ApoA-I and AAT in one or more processing steps.27. The method according to claim 26 , wherein the starting human plasma fraction is selected from one or more of Cohn fractions IV claim 26 , precipitates from Kistler-Nitschmann supernatants A and A+I claim 26 , and ammonium sulfate precipitates.28. The method according to claim 27 , wherein the one or more Cohn fractions IV is Cohn fraction IV.29. The method according to claim 26 , wherein AAT and ApoA-I are not exposed to a pH of 13.69 or above.30. The method according to claim 29 , wherein AAT and ApoA-I are not exposed to a pH ...

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07-11-2013 дата публикации

Downstream bioprocessing device

Номер: US20130296538A1
Автор: Sarfaraz K. Niazi
Принадлежит: Therapeutic Proteins International LLC

Large-scale downstream processing of secreted recombinant proteins is provided in a single device, wherein the contents of a plurality of bioreactors are combined simultaneous to their harvesting and purification resulting in significant savings of time and the cost of manufacturing.

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14-11-2013 дата публикации

VANCOMYCIN B HYDROCHLORIDE CRYSTALLINE FORM 1

Номер: US20130303727A1
Принадлежит:

Vancomycin B Hydrochloride Crystalline Form 1, compositions containing it and methods of prevention or treatment of bacterial infections using it are disclosed. 1providing a mixture comprising Vancomycin B hydrochloride and one volume of water, wherein the vancomycin B hydrochloride is completely soluble in the water;causing Vancomycin B Hydrochloride Crystalline Form 1 to exist in the mixture by adding 1.5 volumes of acetone to the mixture; andisolating the Vancomycin B Hydrochloride Crystalline Form 1.. A process for making Vancomycin B Hydrochloride Crystalline Form 1 comprising: This application claims priority to U.S. patent application Ser. No. 60/882,656, filed Dec. 29, 2006 and is incorporated herein by reference in its entirety.This invention pertains to Vancomycin B Hydrochloride Crystalline Form 1, ways to make it, compositions containing it and methods of treatment of diseases using it.Vancomycin B hydrochloride is a salt of Vancomycin B (CHClNO) that is produced by the actinomycete (formerly ) found in Indonesian and Indian soil.Because the relationship between different crystalline forms of Vancomycin B Hydrochloride may provide guidance for further development, there is an existing need in the chemical and therapeutic arts for identification of different crystalline forms of Vancomycin B Hydrochloride and ways of reproducibly making it.One embodiment of this invention pertains to Vancomycin B Hydrochloride Crystalline Form 1.Still another embodiment of this invention pertains to Vancomycin B Hydrochloride Crystalline Form 1 having substantial crystalline purity.Still another embodiment pertains to compositions made with or comprising an excipient and Vancomycin B Hydrochloride Crystalline Form 1.Still another embodiment pertains to methods for treating bacterial infection in a human comprising administering thereto a therapeutically effective amount of Vancomycin B Hydrochloride Crystalline Form 1.Still another embodiment pertains to a process for ...

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28-11-2013 дата публикации

Purification of Proteins

Номер: US20130317204A1
Автор: Jaber Jad, Moya Wilson
Принадлежит: EMD MILLIPORE CORPORATION

The present invention relates to a selectively soluble polymer capable of binding to a desired molecules in an unclarified mixture containing various biological materials and the methods of using such a polymer to purify a molecule from such a mixture. The polymer is soluble in the mixture under a certain set of process conditions such as pH or temperature and/or salt concentration and is rendered insoluble and precipitates out of solution upon a change in the process conditions. The polymer is capable of binding to the desired molecule (protein, polypeptide, etc) and remains capable of binding to that molecule even after the polymer is precipitated out of solution. The precipitate can then be filtered out from the remainder of the stream and the desired biomolecule is recovered such as by elution and further processed. 1) A method for purifying a biomolecule from an unclarified mixture containing impurities comprising:a. providing the mixture at a set of conditions,b. adding one or more polymers, soluble in said mixture under the set of conditions and capable of reversibly and selectively binding to at least the biomolecule, said one or more polymers being selected from the group consisting of poly(N-isopropylacrylamide), functionalized agarose, polyethylene oxide, cationic polyelectrolytes and anionic polyelectrolytes;c. mixing the one or more solubilized polymers throughout the mixture;d. precipitating the one or more polymers and at least the bound biomolecule out of solution by changing the set of conditions in the mixture; ande. separating the precipitated polymer and at least the bound biomolecule from the mixture.2) The method of wherein the one or more polymers are precipitated by a change in temperature and are selected from the group consisting of poly(N-isopropylacrylamide) claim 1 , functionalized agarose claim 1 , hydroxyalkylcellulose and polyethylene oxide.3) The method of wherein the one or more polymers are precipitated by a change in pH and are ...

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05-12-2013 дата публикации

Methods, compositions, and kits for protein crystallization

Номер: US20130319863A1
Принадлежит: Life Technologies Corp

The present invention provides methods, compositions, and kits for protein crystallization. The present invention involves electrophoretically focusing at least a first protein species within a matrix comprising at least 2 regions of different pH, the protein being present in amount sufficient to permit crystallization within said pH gradient.

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19-12-2013 дата публикации

COMPOSITIONS FOR SEPARATION METHODS

Номер: US20130337528A1
Принадлежит:

This invention relates generally to the fields of separation and conversion technologies, and more particularly to materials for use in tangential-flow filtration techniques. The tangential-flow materials are useful in a wide range of separation and conversion processes, including those reliant on reverse osmosis, microfiltration, ultrafiltration, or nanofiltration semipermeable filtration membranes, and provide efficient methods for purifying or producing various target substances, including biopolymer particles for use in tangential-flow filtration. 152.-. (canceled)53. A method for preparing one or more target substances from a source material , the method comprising contacting the source material with a population of amorphous polymer particles for a time sufficient to allow the amorphous polymer particles to bind one or more target substances or one or more precursors of a target substance or one or more contaminants , separating by tangential flow filtration the one or more contaminants from the particle-bound target substance or precursor thereof or the one or more target substances or precursor thereof from a particle-bound contaminant , and recovering the target substance.54. The method of claim 53 , wherein the population of amorphous polymer particles is a heterogeneous population.55. The method of claim 53 , wherein one or more of the amorphous polymer particles comprises one or more biopolymers selected from a polyester claim 53 , a polythioester or a polyhydroxyalkanoate.56. The method of claim 55 , wherein one or more of the polymer particles comprises a biopolymer selected from a polyester claim 55 , a polythioester or a polyhydroxyalkanoate andi. a polymer particle-forming polypeptide, orii. a polymer particle-binding polypeptide;iii. a polypeptide fusion partner;iv. an affinity ligand;v. an enzyme;vi. a fusion polypeptide comprising two or more of the above; orvii. any combination of any two or more of (i) to (vi) above.57. The method of claim 53 , ...

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26-12-2013 дата публикации

Devices and Methods for Integrated Continuous Manufacturing of Biological Molecules

Номер: US20130345402A1
Принадлежит: Bayer HealthCare LLC

The present invention relates to a process and apparatus for purifying a molecule of interest from a heterogeneous clarified fluid mixture. The apparatus of the invention generally comprises a continuous perfusion fermentation system, a continuous particle removal system integrated with the perfusion fermentation system; and a continuous purification system integrated with the particle removal system, which is maintained under sterile conditions. The process comprises filtering a heterogeneous clarified fluid mixture by continuous ultrafiltration at a specific flow rate below the transition point of the molecule of interest in the pressure-dependent region of the flux versus TMP curve, wherein the specific flow rate is maintained substantially constant throughout the continuous ultrafiltration. 1. A process for purifying a molecule of interest from a heterogeneous clarified fluid mixture , comprising: filtering a heterogeneous clarified fluid mixture by continuous ultrafiltration at a specific flow rate below the transition point of the molecule of interest in the pressure-dependent region of the flux versus TMP curve , wherein the specific flow rate is maintained substantially constant throughout the continuous ultrafiltration.2. The process according to claim 1 , further comprising filtering the clarified mixture at a specific flow rate that produces a wall concentration less than about 20% greater than the retentate concentration.3. The process according to claim 1 , further comprising filtering the clarified mixture at a specific flow rate that produces a wall concentration less than about 15% greater than the retentate concentration.4. The process according to claim 1 , further comprising filtering the clarified mixture at a specific flow rate that produces a wall concentration less than about 10% greater than the retentate concentration.5. The process according to claim 1 , further comprising:filtering the clarified fluid mixture through an ultrafiltration ...

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26-12-2013 дата публикации

EFFICIENT PEPTIDE COUPLINGS AND THEIR USE IN THE SYNTHESIS AND ISOLATION OF A CYCLOPENTA (G) QUINAZOLINE TRISODIUM SALT

Номер: US20130345423A1
Принадлежит: Onyx Pharmaceutical, Inc.

A new method for the synthesis of L-Glutamyl-γ-D-Glutamic acid and its use in the synthesis of (2R)-((4S)-carboxy-4-(4,N-(((6S)-2-(hydroxymethyl)-4-oxo-3,4,7,8-tetrahydro-3H-cyclopenta[g]quinazolin-6-yl)-N-(prop-2-ynyl)amino)benzamido)butanamido)pentanedioic acid, 1 are provided. Also provided is an efficient method for the isolation and purification of the trisodium salt of the abovementioned acid, 2, in a form suitable for long term storage and use in a parenteral dosing form. 1. A method for the synthesis of a L-Glu-γ-D-Glu dipeptide comprising the steps of:a) activating the γ-carboxylic acid of an N-αO-diprotected L-Glu derivative;b) silylating D-glutamic acid; andc) reacting the activated carboxylic acid derivative produced in step a) with the silylated product of step b) to give a protected L-Glu-γ-D-Glu species.2. A method according to claim 1 , wherein in step a) claim 1 , activation comprises conversion of the N-αO-diprotected L-Glu derivative into an activated derivative.3. A method according to claim 2 , wherein activation comprises treatment of the N-αO-diprotected L-Glu derivative with an alkyl chloroformate and a tertiary amine base.4. A method according to claim 3 , wherein the chloroformate is isobutyl chloroformate.5. A method according to whereby the N-αO-diprotected L-Glu derivative is added slowly to a low temperature solution of the alkyl chloroformate and tertiary amine base.6. A method according to claim 5 , wherein the solution of the alkyl chloroformate and tertiary amine base is at a temperature of from −10° C. to −50° C.7. A method according to claim 3 , wherein the tertiary amine base is N-methylmorpholine.8. A method according to wherein the N-αO-diprotected L-Glu derivative is N-benzyloxycarbonyl-L-glutamic acid-α-benzyl ester.9. A method according to wherein in step b) claim 1 , silylation comprises treatment of D-glutamic acid with a silylating agent.10. A method according to claim 9 , wherein the silylating agent is O claim 9 ,N-bis ...

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02-01-2014 дата публикации

LIGAND GRAFT FUNCTIONALIZED SUBSTRATES

Номер: US20140004595A1
Принадлежит: 3M INNOVATIVE PROPERTIES COMPANY

Polyethyleneimine and polyalkylene biguanide ligand functionalized substrates, methods of making ligand functionalized substrates, and methods of using functionalized substrates are disclosed. 1. A filter element comprising one or more layers of a porous , ligand functionalized substrate and extending from the surfaces thereof free-radically grafted biguanide ligand groups to form a free-radically-grafted substrate , wherein the free-radically grafted substrate is of the formula:{'sub': ligand', 'm', 'ligand, 'Substrate˜(M), where Substrate represents the thermoplastic substrate, Mrepresents a polymerized grafted ethylenically unsaturated monomer having pendant biguanide ligand groups, and m is at least two.'}4. The article of claim 1 , wherein the porous substrate is selected from a porous membrane claim 1 , porous non-woven web claim 1 , or porous fiber.5. The article of further comprising free-radically grafted ionic or hydrophilic groups.6. The article of claim 5 , further comprising free-radically grafted quaternary ammonium groups on the surface of the substrate.7. The filter element of comprising at least two layers of the porous claim 1 , ligand functionalized substrate.8. The filter element of wherein the individual layers have different porosity9. The filter element of wherein the individual layers have different degrees of grafting by the grafting monomers.10. The filter element of further comprising an upstream prefilter layer and/or a downstream support layer.11. The filter element of wherein the individual filter elements are pleated.12. The filter element of wherein the pore size of the porous substrate is from 0.1 to 10 μm13. A filter cartridge comprising the filter element of .14. A method of filtration comprising the steps of:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'a) providing the filter element comprising one of more layers of the ligand functionalized base substrate of , and'}b) allowing a moving biological solution containing a ...

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23-01-2014 дата публикации

Binding of pathological forms of proteins using conjugated polyelectrolytes

Номер: US20140024813A1
Принадлежит: BioChromix AB

A method for separation of an aggregated misfolded protein from an environment including a non-aggregating normal form of the protein includes contacting both the misfolded and normal protein with a conjugated polyelectrolyte (CPE) and separating the CPE/protein complex from the other constituents of the sample.

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30-01-2014 дата публикации

Purification of cell culture derived alpha1 protease inhibitor

Номер: US20140031532A1
Принадлежит: Grifols SA

Described herein are methods for purifying recombinant, cell culture derived alpha 1 -protease inhibitor and removing a colored species that co-purifies with the recA1PI protein. Also described are methods for reducing the iron in cell culture derived alpha 1 -protease inhibitor.

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13-02-2014 дата публикации

Method for preparing a depleted plasma material consisting of one or more thrombogenic factors

Номер: US20140044742A1
Принадлежит: LFB SA

The invention concerns a method for preparing a plasma product depleted of one or more thrombogenic factors, comprising the combination of at least two steps chosen from among an ethanol fractionation step, a filtration-adsorption step, a precipitation step with caprylic acid and a chromatography step on ion exchange resin.

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13-02-2014 дата публикации

DRUG SUBSTANCES, PHARMACEUTICAL COMPOSITIONS AND METHODS FOR PREPARING THE SAME

Номер: US20140044759A1
Принадлежит:

Drug substances, which comprise a solid amorphous forms of a compound of structural formula I and have a BET specific surface area of up to about 94 m/g, pharmaceutical compositions comprising such drug substances, processes for preparing such drug substances and uses of such drug substances and pharmaceutical compositions are disclosed. 2. The drug substance according to claim 1 , wherein said drug substance has a BET specific surface area of from about 2.9 m/g to about 9.6 m/g.3. The drug substance according to claim 1 , wherein said drug substance has a BET specific surface area of from about 2.9 m/g to about 9.4 m/g.4. A pharmaceutical composition comprising at least one drug substance according to and at least one pharmaceutically acceptable carrier.5. The pharmaceutical composition according to claim 4 , further comprising at least one excipient.7. The process according to claim 6 , wherein said distilling step b) is conducted.8. The process according to claim 7 , wherein said distilling step b) is conducted at a temperature in a range from about −15.0° C. to about 35.0° C.9. The process according to claim 7 , wherein said distilling step b) is conducted at a temperature in a range from about 15.0° C. to about 30.1° C.10. The process according to claim 7 , wherein said distilling step b) is conducted at a temperature in a range from about 15.0° C. to about 24.6° C.11. The process according to claim 10 , wherein said distilling step b) is conducted at a temperature in a range from about 15.1° C. to about 24.6° C. for the first 10 hours of the distilling.12. The process according to claim 7 , wherein said distilling step b) is conducted at a temperature in a range from about −15.0° C. to about 15.0° C.13. The process according to claim 7 , wherein said distilling step b) is conducted over a total of 20 to 30 hours.14. The process according to claim 6 , wherein said filtering step c) is conducted at a temperature in a range from about −15.0° C. to about 15.0° C. ...

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06-03-2014 дата публикации

ISOLATED EGG PROTEIN AND EGG LIPID MATERIALS, AND METHODS FOR PRODUCING THE SAME

Номер: US20140066596A1
Автор: Mason David
Принадлежит:

A method for separating proteins and fats from an egg mixture is disclosed herein. The method includes a step of microfiltration of the egg mixture, wherein microfiltration includes pumping across a filter an egg mixture containing egg yolk and egg whites (albumen). An egg powder obtained from egg and a high gel strength egg powder are also disclosed. 1. A method for separating proteins from an egg mixture , the method comprising:(a) obtaining an egg mixture comprising egg-derived lipids and egg-derived proteins; and(b) microfiltration of the egg mixture to obtain an isolated protein composition;wherein the microfiltration comprises pumping the egg mixture across a membrane with a pore size less than 0.40 microns at a pressure of less than 60 psi.2. The method for separating proteins from an egg mixture of claim 1 , wherein the membrane comprises polyvinylidene fluoride.3. The method of claim 1 , wherein microfiltration comprises pumping the egg mixture across a spiral wound membrane.4. The method of claim 1 , wherein microfiltration comprises pumping the egg mixture across a filter with a pore size of equal to or less than 0.30 microns5. The method of claim 1 , wherein microfiltration comprises pumping the egg mixture across a filter with a pore size of equal to or less than 0.20 microns at a pressure of less than about 30 PSI.6. The method for separating proteins from an egg mixture of claim 1 , wherein the egg mixture comprises at least 1 percent egg-derived lipid.7. The method for separating proteins from an egg mixture of claim 1 , wherein the egg mixture comprises at least 5 percent egg-derived lipid.8. The method of claim 1 , wherein the membrane is constructed from a hydrophilic material.9. The method of claim 1 , wherein the pH of the mixture is not manipulated during separation.10. The method of claim 1 , wherein the egg mixture comprises inedible egg.11. The method of claim 1 , wherein the egg mixture comprises edible egg.12. The method of claim 1 , ...

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06-03-2014 дата публикации

Purification of biological conjugates by size exclusion chromatography

Номер: US20140066605A1
Принадлежит: ABBOTT LABORATORIES

A method for separating a biological conjugate from an aggregate. The molecular weight of the biological conjugate ranges from about 10 kDa to about 1000 kDa. In one embodiment, the method comprises the steps of: (e) providing a mixture comprising the biological conjugate and the aggregate, wherein the biological conjugate has a molecular weight of from about 10 kDa to about 1000 kDa; (f) providing a chromatography column containing a gel, wherein the gel comprises at least one polysaccharide; (g) introducing the mixture of step (a) into the chromatography column; (h) recovering the biological conjugate from the column.

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20-03-2014 дата публикации

TUMOUR MARKER PROTEINS AND USES THEREOF

Номер: US20140080736A1
Принадлежит: Onclmmune Limited

Tumour marker proteins and their preparation from fluids from one or more cancer patients, wherein said fluids are those which collect in a body cavity or space which is naturally occurring or which is the result of cancer or medical intervention for cancer. The present application also relates to preparation of tumour marker proteins from excretions taken from patients with cancer. The tumour marker proteins are useful as immunoassay reagents in the detection of cancer-associated anti-tumour marker autoantibodies. 1. A method of detecting cancer-associated anti-tumor autoantibodies in a sample from an individual , comprising:contacting the sample with an immunoassay reagent; and detecting a presence of complexes formed by specific binding of the immunoassay reagent to any cancer-associated anti-tumor autoantibodies present in the sample,wherein the immunoassay reagent comprises two or more tumor marker proteins, one or more of which having been prepared from a tumor-induced bodily fluid produced in a body cavity or space in the presence of the tumor of one or more cancer patients, wherein the bodily fluid contains more cancer-associated forms of the tumor marker protein than a non-tumor-induced bodily fluid in the same patient, and the bodily fluid is not a fluid from the systemic circulation,wherein the one or more tumor marker proteins prepared from a bodily fluid exhibit selective reactivity with cancer-associated anti-tumor autoantibodies, wherein the tumor marker proteins are over-expressed or altered forms of wild-type proteins, andwherein detection of complexes indicates the presence of cancer-associated anti-tumor autoantibodies in the individual.2. The method of claim 1 , further comprising detecting and/or quantitatively measuring the presence of two or more types of autoantibodies claim 1 , wherein each one of the two or more types of the autoantibodies is immunologically specific to a different tumor marker protein or to different epitopes of the same ...

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27-03-2014 дата публикации

METHOD OF MAKING LIGAND FUNCTIONALIZED SUBSTRATES

Номер: US20140087445A1
Принадлежит: 3M INNOVATIVE PROPERTIES COMPANY

Ligand functionalized substrates, methods of making ligand functionalized substrates, and methods of using functionalized substrates are disclosed. 2. The ligand-functional substrate of wherein said crosslinking monomers (c) have two or more free-radically polymerizable groups.3. The method of wherein said crosslinking monomer c) is of the formula:{'br': None, 'sub': 2', 'a', 'b, 'sup': 1', '10, '[CH═CH—C(O)—X]—R-Q-Z,'}where Z is an acryloyl or non-acryloyl, ethylenically unsaturated polymerizable group,{'sup': 1', '3', '3, 'sub': 1', '4, 'Xis —O— or —NR, where Ris H or C-Calkyl,'}{'sup': '1', 'sub': '2', 'Q is a divalent linking group selected from a covalent bond “—”, —O—, —NR—, —CO— and'}{'sup': 1', '1, 'sub': 1', '4, '—CONR—, where Ris H or C-Calkyl; and'}{'sup': '10', 'Ris an alkylene group of valence a+b, and optionally containing one or more catenary oxygen atoms and/or one or more hydroxyl groups;'}and a and b are each at least one.4. The ligand-functional substrate of wherein the crosslinking monomer c) comprises a poly(alkylene oxide) compound having at least one acryloyl group and at least one additional ethylenically unsaturated claim 2 , free-radically polymerizable group.5. The ligand-functional substrate of claim 2 , wherein the monomer c) is of the formula:{'br': None, 'sup': '1', 'sub': 2', 'n', '2, 'Z-Q-(CH(R)—CH—O)—C(O)—CH═CH,'}wherein Z is an acryloyl or non-acryloyl, polymerizable ethylenically unsaturated group,{'sup': 1', '1', '1', '1, 'sub': 1', '4', '2', '1', '4, 'Ris a H or a Cto Calkyl group, and n is from 2 to 100, preferably 5 to 20, and Q is a divalent linking group selected from a covalent bond “—”, —O—, —NR—, —CO— and —CONR—, where Ris H or C-Calkyl.'}6. The ligand-functional substrate of further comprising one or more additional monomers having a free-radically polymerizable group and a hydrophilic group.7. The ligand functionalized substrate of wherein said hydrophilic monomers are of the formula:{'br': None, 'sub': 2', '2', 'n, ' ...

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03-04-2014 дата публикации

Methods for purification of alpha-1-antitrypsin and apolipoprotein a-1

Номер: US20140094591A1
Принадлежит: CSL Behring GmbH Deutschland

This invention relates to protein separation and purification methods for both alpha-1-antitrypsin (AAT, also known as alpha-1 proteinase inhibitor, API, and A.sub.1-PI) and Apolipoprotein A-I (ApoA-1) from, for example, a fraction of human blood plasma. In certain embodiments, the invention provides methods for separating AAT from ApoA-1 at the initial stage of purification, so that the same starting material can be used as a source for both proteins. The methods further pertain to providing compositions of AAT and of ApoA-1 suitable for pharmaceutical use and are suitable for large-scale purification.

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03-01-2019 дата публикации

PRODUCT ANALOGS OR COMPONENTS OF SUCH ANALOGS AND PROCESSES FOR MAKING SAME

Номер: US20190000112A1
Принадлежит:

Provided are food products that are derived from non-animal sources that have one or more of the following: color, taste, nutritional content, and other qualities similar to those of dairy products and/or other types of food products. Also provided are processes for production of such dairy-like food products and/or other types of food product analogs. 1. A method for obtaining a yield of refined protein component from a non-animal natural and/or modified non-animal natural source comprising the steps of:a) obtaining a protein preparation from the non-animal natural or modified non-animal natural source;b) washing the protein preparation at a wash pH;c) extracting the protein preparation at an extraction pH to obtain an aqueous protein solution;d) separating the aqueous protein solution from non-aqueous components;e) adding salt;f) precipitating the protein from the aqueous protein solution at a precipitation pH to obtain a protein precipitate;g) separating the protein precipitate from non-precipitated components; andh) washing the protein precipitate to obtain the refined protein component at the substantial yield.2. The method of claim 1 , wherein the non-animal natural or modified non-animal natural source is a plant.3. The method of claim 1 , wherein the plant is a legume.4. The method of claim 1 , wherein the legume is pea.5. The method of claim 1 , wherein the wash pH is between about 3 and about 5.6. The method of claim 1 , wherein the extraction pH is between about 8 and about 9.7. The method of claim 1 , wherein the salt added is calcium dichloride at a concentration of between about 50 mM and about 100 mM.8. The method of claim 1 , wherein the precipitation pH is between about 5 and about 6.9. The method of claim 1 , wherein the method further comprises the step of adding between about 0.05% and about 12% by weight of polysaccharide to the aqueous protein solution.10. The method of claim 1 , wherein the yield is between about 50% and about 90%.11. A ...

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04-01-2018 дата публикации

METHOD FOR OBTAINING A PEPTIDE ISOLATE FROM A BIOMASS OF PROTEIN-ENRICHED MICROALGAE

Номер: US20180000116A1
Принадлежит: ROQUETTE FRERES

The invention relates to a peptide isolate isolated from a biomass of protein-rich microalgae, characterized in that it comprises: 1. A peptide isolate isolated from a biomass of protein-rich microalgae , said isolate comprising:soluble peptides with a molecular weight of between 1 and 20 kDa,a protein content expressed as N.6.25 of more than 95%, wherein total amino acid content of said protein substantially comprises arginine and glutamic acid.2Chlorella.. The isolate according to claim 1 , wherein the microalgae are of genus3. The isolate according to claim 1 , wherein the content of arginine and glutamic acid is more than 80% by weight expressed relative to the total amino acids.4. The isolate according to claim 1 , wherein the content of arginine and glutamic acid is respectively at least 40% exppresse relative to the total amino acid content.5. The isolate according to claim 4 , wherein the content of arginine is between 47 and 48% for arginine and between 49 to 50% for glutamic acid expressed relative to the total amino acid content.6Chlorella. The isolate according to claim 1 , prepared by fermentation of a biomass of protein-rich microalgae genus method which comprises:{'i': 'Chlorella', 'a “batch” fermentation phase in a fermenter of the biomass of protein-rich microalgae genus after seeding the fermenter, by supplying in a single portion an amount of glucose of between 15 and 25 g/l,'}{'sub': 3', '3, 'an exponential fed batch phase during which glucose is supplied gradually and pH regulation is performed initially by a NH/KOH mixture and subsequently replaced by regulation with NHalone.'}7Chlorella. The isolate according to claim 1 , obtained from a biomass of protein-rich microalgae of genus claim 1 , by a method which comprises:optionally, washing the biomass so as to remove the interstitial soluble compounds,thermal permeabilization of the biomass at a temperature of between 50 and 150° C., for a time of between about 10 seconds and about 5 minutes, ...

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02-01-2020 дата публикации

Load bearing crowded collagen constructs

Номер: US20200000962A1
Автор: Hamed Alavi, Ya Guo
Принадлежит: MEDTRONIC INC, Medtronic Vascular Inc

Load bearing crowded collagen material for use in engineered tissue may be formed by crowding collagen fibrils. The crowding may be performed by dialysis. The fibrils may be formed by neutralizing an acidic solution comprising collagen monomers.

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07-01-2016 дата публикации

WHEY PROTEIN CONCENTRATE, ITS PREPARATION AND ITS USE

Номер: US20160002288A1
Принадлежит: FRIESLANDCAMPINA NEDERLAND HOLDING B.V.

The invention pertains to process for manufacturing whey protein concentrate (WPC) from whey, said process involving (a) providing acidified whey; (b) increasing the pH of said acidified whey using one or more carbonate salt(s), preceded and/or followed by ultrafiltration, and (c) subjecting the ultrafiltered carbonate-containing whey to spray drying. A WPC is provided having improved functional properties, particularly increased gel strength and reduced salt sensitivity (i.e. meaning that the functional properties of the WPC are affected by salt to a lesser extent). 114.-. (canceled)15. A process for manufacturing whey protein concentrate (WPC) from whey , the process comprising:(a) increasing the pH of acidified whey using one or more carbonate salt(s),(b) subjecting the acidified whey to ultrafiltration before and/or after increasing the pH, and(c) subjecting the ultrafiltered whey to spray drying.16. The process according to claim 15 , wherein the pH is increased to at least 6.3.17. The process according to claim 16 , wherein the pH is increased to between 6.4-7.0.18. The process according to claim 15 , wherein the pH of the whey after spray drying is higher than 7.019. The process according to claim 18 , wherein the pH of the whey after spray drying is between 7.0 and 8.5.20. The process according to claim 15 , wherein the retentate obtained after ultrafiltration is subjected to diafiltration.21. A whey protein concentrate (WPC) having (i) a whey protein content of at least 70% claim 15 , based on dry matter claim 15 , (ii) a pH of at least 6.6 claim 15 , and (iii) a total carbonate content in the concentrate of 0.7-1.4%.22. The WPC according to claim 21 , having a pH of at least 7.0.23. The WPC according to in the form of a powder.24. The WPC according to claim 21 , having a pH between 7.0 and 8.5.25. The WPC according to claim 21 , in the form of a gel claim 21 , wherein the gel has a gel strength of at least 6000 grams claim 21 , by measuring the maximum ...

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07-01-2016 дата публикации

METHOD OF PURIFYING AN ANTIBODY

Номер: US20160002289A1
Принадлежит: Synthon Biopharmaceuticals B.V.

A method of purifying an antibody composition comprises application of anion exchange chromatography late in the purification process. An ultrafiltration/diafiltration-purified antibody composition is subjected to anion exchange chromatography (AEX) to form a pharmaceutically-pure antibody composition. 1. A method of purifying an antibody composition , which comprises subjecting a UF/DF-purified antibody composition to anion exchange chromatography (AEX) to form a pharmaceutically pure antibody composition.2. The method of claim 1 , wherein said UF/DF-purified antibody composition has an antibody concentration of at least 1 mg/ml.3. The method of claim 2 , wherein said UF/DF-purified antibody composition has an antibody concentration of at least 10 mg/ml.4. The method of claim 3 , wherein said UF/DF-purified antibody composition has an antibody concentration of not greater than 250 mg/ml.5. The method of claim 4 , wherein said UF/DF-purified antibody composition has an antibody concentration of 20 to 50 mg/ml.6. The method of claim 1 , which further comprises filling said pharmaceutically pure antibody composition into vials.7. The method according to claim 6 , which further comprises aseptically filtering said pharmaceutically pure antibody composition prior to said filling step.8. The method according to claim 7 , which further comprises at least one of the following steps prior to said filling step: (a) adding an excipient to said pharmaceutically-pure antibody composition; (b) concentrating said pharmaceutically-pure antibody composition; (c) diluting said pharmaceutically-pure antibody composition; and/or (d) adjusting the pH of said pharmaceutically-pure antibody composition.9. The method of claim 1 , which further comprises subjecting a partially-purified antibody composition to UF/DF to form said UF/DF-purified antibody composition.10. The method of claim 9 , which further comprises subjecting an antibody cell culture harvest to an antibody capture step and ...

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07-01-2016 дата публикации

MATERIALS AND METHODS FOR REMOVING ENDOTOXINS FROM PROTEIN PREPARATIONS

Номер: US20160002290A1
Принадлежит:

A method includes (i) adding allantoin in a supersaturating amount to a protein preparation including a desired protein and at least one endotoxin as a contaminant, (ii) removing solids after the adding step to provide a sample for further purification by void exclusion chromatography on a packed particle bed of electropositive particles in a column, the packed particle bed having an interparticle volume, (iii) applying a sample volume to the packed particle bed, wherein the electropositive particles support void exclusion chromatography, and wherein the sample volume is not greater than the interparticle volume, and (iv) eluting a purified sample including the desired protein and a reduced amount of the endotoxin. The method is optionally carried out with only the allantoin treatment or only the void exclusion chromatography. 116.-. (canceled)17. A method comprising: (i) adding allantoin in a supersaturating amount to a protein preparation comprising a desired protein and an amount of at least one endotoxin as a contaminant; (ii) removing solids from the protein preparation to provide a sample for further purification by void exclusion chromatography using a packed particle bed of electropositive particles in a column , the packed particle bed having an interparticle volume; (iii) applying a sample volume of the sample to the packed particle bed , wherein the electropositive particles support void exclusion chromatography , and wherein the sample volume is not greater than the interparticle volume , and (iv) eluting a purified sample comprising the desired protein and a reduced amount of the at least one endotoxin , wherein the eluted desired protein resides in a buffer to which the column was equilibrated , independently from a buffer content of the sample applied to the column.18. The method of claim 17 , wherein the supersaturating amount of allantoin comprises an amount selected from the group consisting of: (i) about 10% claim 17 , (ii) about 5% claim 17 , (ii ...

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03-01-2019 дата публикации

NON-CHROMATOGRAPHIC SEPARATION OF POLYPEPTIDES USING THE COMBINATION OF A STERIC EXCLUSION AGENT AND A CHARGE NEUTRALIZATION AGENT

Номер: US20190002495A1
Принадлежит: KBI Biopharma, Inc.

The present invention relates to methods of separating polypeptides of interest from contaminants, including DNA and undesired polypeptides, in a common fluid by precipitation of the polypeptides of interest or, alternatively, the contaminants using the combination of steric exclusion agents and charge neutralization agents. 1. A method of separating a polypeptide of interest and a contaminant in a fluid by precipitation , comprising contacting the fluid with a combination of a steric exclusion agent and a charge neutralization agent in a single precipitation step , thereby separating the polypeptide of interest from the contaminant by precipitation.2. The method of claim 1 , wherein the fluid is contacted with a single solution comprising the steric exclusion agent and the charge neutralization agent.3. The method of claim 1 , wherein the fluid is contacted with a first solution comprising the steric exclusion agent and a second solution comprising the charge neutralization agent.4. The method of claim 1 , wherein the contaminant is precipitated and the polypeptide of interest remains in solution.5. The method of claim 1 , wherein the polypeptide of interest is precipitated and the contaminant remains in solution.6. The method of claim 1 , wherein the polypeptide of interest is a recombinant polypeptide produced in a cell culture.7. The method of claim 6 , wherein the cell culture is a mammalian cell culture.8. The method of claim 6 , wherein the cell culture is a bacterial cell culture.9. The method of claim 6 , wherein the cell culture is a fungal cell culture.10. The method of claim 1 , wherein the fluid is harvested cell culture medium.11. The method of claim 1 , wherein the polypeptide of interest is an antibody or antibody fragment.12. The method of claim 1 , wherein the polypeptide of interest is an enzyme.13. The method of claim 1 , wherein the steric exclusion agent is selected from the group consisting of dextran claim 1 , starch claim 1 , and ...

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04-01-2018 дата публикации

Non-Denatured Proteins Derived From a Biomass Source

Номер: US20180002374A1
Автор: Laubsch Kenneth L.
Принадлежит:

A biomass-derived protein compound has a high concentration of protein and can be made to have a very low concentration of fat and water; even when the biomass feedstock has a high fat concentration. The biomass-derived protein compound may be a whole protein that is non-denatured and enzymatically digestible. This unique protein compound can be produced from molecules from more than one source organism, including various animals and/or plant feedstocks. The unique protein compound is derived from a unique biomass method and apparatus for the treatment of a biomass stream to extract and separate an essentially solvent-free product from the biomass stream. In this unique method the solids content of the biomass stream is increased by bringing the biomass stream into contact with a moderately pressurized liquefied gas solvent, to create a high solids content biomass stream and introducing the high solids content biomass stream to an extraction apparatus. 1. A biomass-derived protein compound comprising:a mixture of protein molecules from more than one source organism wherein the mixture contains at least 65% whole, non-denatured protein molecules by weight, less than 15% fat by weight, and less than 10% water by weight.2. The biomass-derived protein compound of claim 1 , wherein at least 75% of the protein molecules in the compound contain each of the following essential amino acids: histidine claim 1 , isoleucine claim 1 , leucine claim 1 , lysine claim 1 , methionine claim 1 , phenylalanine claim 1 , threonine claim 1 , tryptophan claim 1 , and valine.3. The biomass-derived protein compound of claim 1 , wherein the biomass-derived protein compound has a neutral pH with a pH between 6 and 8.4. The biomass-derived protein compound of claim 1 , wherein a substantial portion of the protein molecules are enzymatically digested.5. The biomass-derived protein compound of claim 1 , wherein the biomass from which the protein compound is derived contains greater than 50% fat ...

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03-01-2019 дата публикации

METHODS OF PURIFICATION AND/OR VIRAL INACTIVATION

Номер: US20190002518A1
Принадлежит: FERRING B.V.

Methods of purification and/or viral deactivation of a protein (e.g. glycoprotein) comprising a step of treating the protein (e.g. glycoprotein) with a combination of caprylic acid and ethanol. 1. A method of purification of a protein , the method comprising a step of treating the protein with a combination of caprylic acid and ethanol.2. A method of viral inactivation in a protein , the method comprising a step of treating the protein with a combination of caprylic acid and ethanol.3. A method according to claim 1 , which comprises treating a solution of the protein with a combination of caprylic acid and ethanol.4. A method according to claim 1 , wherein the protein is a glycoprotein.5. A method according to claim 1 , wherein the protein is FSH claim 1 , hCG or LH.6. A method according to claim 1 , wherein the protein is a recombinant glycoprotein.7. A method according to claim 1 , wherein the protein is a recombinant protein produced in a cell by a method comprising culturing the cell in a suitable medium and harvesting the recombinant protein from said cell and/or said medium.8. A method according to claim 7 , wherein the cell is a mammalian cell.9. A method according to claim 1 , wherein treating the protein with a combination of caprylic acid and ethanol takes place at pH 2 to pH 6.5.10. A method according to claim 1 , wherein the caprylic acid concentration is from 10 mM to 30 mM caprylic acid.11. A method according to which comprises:(a) treating the protein with ethanol and caprylic acid for an incubation time of from 1 minute to 6 hours at a temperature of 23±2° C. with stirring;(b) treating the protein with ethanol and caprylic acid at a temperature of 4°-8° C. for an incubation time of from 1 minute to 32 hours, without stirring;(c) treating the protein with ethanol and caprylic acid for an incubation time of from 0.5 hours to 1 hour at a temperature of 23±2° C. with stirring, followed by reducing the temperature to a temperature of 4°-8° C. and ...

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07-01-2021 дата публикации

METHOD FOR REMOVING N-TERMINAL TRUNCATED AND ABNORMAL VARIANTS IN RHNGF

Номер: US20210002341A1
Принадлежит:

A method for removing an N-terminal truncated variant and an abnormal variant in recombinant human nerve growth factor (rhNGF) is provided. An rhNGF raw material loaded on a cation-exchange material is washed with a washing liquid to obtain a washed raw material from which an N-terminal truncated variant and an abnormal variant have been removed, where the washing liquid has higher electrical conductivity than the rhNGF raw material. Cation-exchange chromatography (CEC) elution is then performed on the washed raw material with an elution buffer having higher electrical conductivity than the washing liquid. A purified rhNGF product is obtained from the eluate. 1. A method for removing an N-terminal truncated variant and an abnormal variant in recombinant human nerve growth factor (rhNGF) , comprising:1) washing with a washing liquid an rhNGF raw material loaded on a cation-exchange material, thereby obtaining a washed raw material from which an N-terminal truncated variant and an abnormal variant have been removed, wherein said washing liquid is a washing buffer having higher electrical conductivity than the rhNGF raw material; and2) performing cation-exchange chromatography (CEC) elution on the washed raw material of step 1) with an elution buffer having higher electrical conductivity than the washing liquid in step 1), and collecting an eluate from which a purified rhNGF product is obtained.2. The method of claim 1 , wherein the electrical conductivity of said washing liquid in step 1) is 20˜30 mS/cm.3. The method of claim 1 , wherein said washing liquid in step 1) is an NaCl-containing buffer with an NaCl content of 200˜300 mM.4. The method of claim 1 , wherein step 1) comprises: loading the cation-exchange material with said rhNGF raw material claim 1 , washing with the washing liquid claim 1 , and discarding a resulting outflowing liquid.5. The method of claim 1 , wherein said rhNGF raw material in step 1) is a preliminarily purified product obtained by ...

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03-01-2019 дата публикации

TRANSGLYCOSYLATION OF ENDO-S AND ENDO-S MUTANTS FOR ANTIBODY GLYCOSYLATION REMODELING

Номер: US20190002542A1
Принадлежит:

The present invention provides for a one-pot enzymatic approach which does not require removal of the enzyme and purification of the intermediate after deglycosylation step, and the Endo-S treatment is able to do both deglycosylation and transglycosylation. The one-pot strategy of the present invention enables chemoenzymatic synthesis of an azido-tagged N-glycoform of monocloncal antibodies which could be further modified through orthogonal chemical ligation for various applications. 2Streptococcus pyogenes. The method of claim 1 , wherein the Endo-S enzyme remains in the single container during the entire process.3. The method of claim 1 , wherein the high-mannose or hybrid glycan oxazoline has a predetermined oligosaccharide component with a defined number and type of sugar residues and with specific linkage types.4. The method of claim 4 , wherein the high mannose oxazoline is selected from the group consisting of penta- claim 4 , hexyl- claim 4 , hepta- claim 4 , octyl- claim 4 , nona- claim 4 , deca- claim 4 , or undeca-saccharide oxazolines.5. The method of claim 4 , wherein the hybrid glycan oxazoline comprises mannose claim 4 , sialic acid claim 4 , D-galactose and/or L-fucose.6. The method of claim 4 , wherein the hybrid glycan oxazoline comprises a Man3 glycan oxazoline protected by the addition of an extra moiety to resist hydrolysis by the wild type Endo-S enzyme.7. A single pot remodeling method of a core fucosylated or nonfucosylated IgG or IgG-Fc fragment with an oligosaccharide having a predetermined oligosaccharide component with a defined number and type of sugar residues and with specific linkage types claim 4 , the method comprising:providing a single potproviding and introducing into the single pot a core fucosylated or nonfucosylated IgG or IgG-Fc fragment obtained from natural or recombinant sources carrying heterogeneous N-glycans;{'i': 'Streptococcus pyogenes', 'treating the natural or recombinant IgG or IgG-Fc fragment with an wild type ...

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04-01-2018 дата публикации

Flex plate with removable inserts and cover

Номер: US20180003599A1
Принадлежит: BROOKHAVEN SCIENCE ASSOCIATES LLC

Technologies are described for methods and systems effective for flex plates. The flex plates may comprise a base plate. The base plate may include walls that define an insert location opening in the base plate. The insert location opening in the base plate may be in communication with a securement area. The flex plates may comprise an insert. The insert may include a reservoir region and a crystallization region separated by a wall including channels. The reservoir region and the crystallization region may include a backing. The insert may further include securement tabs. The securement tabs may be configured to secure the insert to the base plate at the securement area.

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07-01-2021 дата публикации

METHODS FOR EXTRACTING PROTEINS FROM BLOOD PLASMA

Номер: US20210003552A1
Принадлежит:

A method of producing protein products including alpha-1-proteinase inhibitor, gamma globulin, albumin, and other proteins from plasma includes steps of: (1) adding a salt to the blood product to produce a first intermediate, wherein the salt comprises between 11-13 wt % of the first intermediate; (2) separating the first intermediate to produce a first supernatant and a first paste; (3) adding a salt to the first intermediate to produce a second intermediate, wherein the salt comprises between 21-23 wt % of the second intermediate; (4) separating the second intermediate to produce a second supernatant and a second paste; (5) separating a third intermediate from the second supernatant by affinity chromatography; and (6) separating the third intermediate by ion exchange chromatography to produce an eluate containing the protein product. Advantageously, the inventive methods are simple and produce alpha-1-proteinase inhibitor, gamma globulin, albumin, and other proteins in high yields. 1. A method of separating A1P1 from a blood plasma containing product , the method comprising:thawing a frozen blood plasma product followed by stirring at a temperature suitable for dissolving a cryoprecipitate to generate the blood plasma containing product;adding a salt to the blood plasma containing product to produce a first intermediate, wherein the salt comprises between 11-20 wt % of the first intermediate;separating the first intermediate to produce a first supernatant and a first paste;adding a salt to the first supernatant to produce a second intermediate, wherein the salt comprises between 15-30 wt % of the second intermediate;separating the second intermediate to produce a second supernatant and a second paste;separating the second supernatant by affinity chromatography using an A1P1-specific affinity media to generate a flow-through fraction and a first eluate, wherein the first eluate comprises A1P1.2. The method of claim 1 , wherein the step of adding the salt to the ...

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13-01-2022 дата публикации

NON-ANTIBODY HIGH-AFFINITY-BASED SAMPLE PREPARATION, SORBENT, DEVICES AND METHODS

Номер: US20220009960A1
Принадлежит:

In various aspects, the present disclosure pertains to sorbents for isolating at least one target protein from a liquid sample, the sorbents comprising a solid support comprising attached at least one attached high affinity reagent with an affinity for the at least one target protein. Other aspects of the present disclosure include kit that contain such sorbents and methods of treating samples using the same. 1. A sorbent for isolating at least one target protein from a liquid sample , the sorbent comprising a solid support comprising attached aptamers or affimers with an affinity for the at least one target protein.2. The sorbent of claim 1 , wherein the aptamers or affimers are linked to the solid support by a chemically cleavable linkage or wherein the aptamers or affimers are linked to the solid support by an enzymatically cleavable linkage.3. The sorbent of claim 2 , wherein the solid support comprises an attached enzyme that is adapted to enzymatically cleave the aptamers or affirmers from the solid support claim 2 , thereby allowing a complex of the aptamer or affirmer and the target protein to be released from the solid support for further processing.4. The sorbent of claim 1 , wherein the solid support comprises an attached enzyme that is adapted to digest the target protein after activation of the attached enzyme.5. The sorbent of claim 4 , wherein the enzyme is attached to the solid support through a flexible linker or wherein the enzyme is attached to the solid support through a cleavable linker.6. The sorbent of claim 4 , wherein the enzyme is attached to the solid support through an enzymatically cleavable linker or wherein the enzyme is attached to the sorbent through a chemically cleavable linker.7. The sorbent of claim 1 , wherein the sorbent is disposed in a multi-well strip claim 1 , a multi-well plate claim 1 , a column claim 1 , a single-use cartridge claim 1 , or an ELISA plate.8. A kit comprising a sorbent in accordance with claim 1 , and one ...

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13-01-2022 дата публикации

Anti-vegf protein compositions and methods for producing the same

Номер: US20220009997A1
Автор: Matthew Franklin
Принадлежит: Regeneron Pharmaceuticals Inc

The present disclosure pertains to compositions comprising anti-VEGF proteins and methods for producing such compositions.

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13-01-2022 дата публикации

ANTI-VEGF PROTEIN COMPOSITIONS AND METHODS FOR PRODUCING THE SAME

Номер: US20220009999A1
Принадлежит:

The present disclosure pertains to compositions comprising anti-VEGF proteins and methods for producing such compositions. 1. A method of producing aflibercept , comprising:(a) binding aflibercept from a clarified harvest cultured in a chemically defined medium (CDM) to a Protein A resin, wherein said aflibercept includes variants that have at least one oxidized amino acid residue selected from the group consisting of tryptophan, histidine, phenylalanine, tyrosine and a combination thereof;(b) eluting said aflibercept of step (a) forming an affinity eluate, wherein said eluate has a first color,wherein said first color is no more yellow-brown than European Color Standard BY1;(c) subjecting said eluate comprising aflibercept to anion exchange chromatography (AEX) column; and(d) collecting a flowthrough fraction, wherein said flowthrough fraction has a second color,wherein said second color has a European Color Standard BY2 or greater, and wherein said first color of said affinity eluate is a more intense yellow brown color than said second color of said flowthrough fraction when said affinity eluate and flowthrough fraction protein concentrations are normalized.2. The method of claim 1 , wherein said cell is selected from a group consisting of CHO claim 1 , NS0 claim 1 , Sp2/0 claim 1 , embryonic kidney cells and BHK.3. The method of claim 1 , wherein said oxidized amino acid residue is histidine.4. The method of claim 1 , wherein said oxidized amino acid residue is tryptophan.5. The method of claim 1 , wherein said AEX column comprises an anionic exchange substituent including diethylaminoethyl (DEAE) claim 1 , quaternary aminoethyl (QAE) and quaternary amine (Q) groups.6. The method of claim 1 , wherein said aflibercept variant is selected from an amino acid residue on a polypeptide having an amino acid sequence as set forth in the group consisting of: SEQ ID NO.: 17 claim 1 , SEQ ID NO.: 18 claim 1 , SEQ ID NO.: 19 claim 1 , SEQ ID NO.: 20 claim 1 , SEQ ID NO.: 21 ...

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03-01-2019 дата публикации

Enzymatic sample purification

Номер: US20190003937A1
Принадлежит: Hewlett Packard Development Co LP

An enzymatic purification method involves the introduction of a sample comprising a target analyte and amino acids into a porous matrix of a reaction chamber. The reaction chamber includes first pores and second pores. The first pores contain polypeptide synthesis enzymes that react with the amino acids to form polypeptides. First pores having a first size to be accessible by amino acids but inaccessible by the subsequently formed polypeptides. The second pores have a second size greater than the first size, are in contact with the first pores and form a series extending from within the reaction chamber to a waste chamber. The formed polypeptides are migrated through the series of second pores to the waste chamber. The target analyte of the sample is extracted from the reaction chamber.

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01-01-2015 дата публикации

SEPARATION OF BIOCOMPONENTS FROM MANURE

Номер: US20150005482A1
Автор: Roa-Espinosa Aicardo
Принадлежит:

A process for separating amino acids and peptides from raw and digested farm manure is disclosed. The raw manure may be recovered from sand bedding used in the cow stalls of some farms. The process involves the steps of precipitating out struvite and separating a stream rich in amino acids and peptides from a mineral rich stream in a dissolved air floatation machine. Struvite precipitation is accomplished by the addition of polydicyandiamide and an acrylate based polymer, while the separation of the stream rich in fibers, amino acids and peptides from a mineral rich streams is accomplished by the addition of an acrylamide chloride copolymer. 1. A substantially continuous process for separating a stream rich in fibers , amino acids and peptides and a stream rich in minerals from raw farm manure , said process comprising the steps of:providing a raw manure stream for treatment, said raw manure stream containing magnesium ammonium phosphate, calcium phosphate, amino acids, peptides fibers and minerals, said minerals containing nitrogen, phosphorus, potassium and sulfur;mechanically removing particles having a predetermined size;precipitating magnesium ammonium phosphate and calcium phosphate, said precipitating being accomplished by the addition of between about 5 parts per million and about 75 parts per million on a dry weight basis of polydicyandiamide polymer, said polydicyandiamide polymer having a weight average molecular weight of between about 3000 and about 150,000; andseparating the stream rich in amino acids, peptides and fibers from the stream rich in minerals, said separating being accomplished in a dissolved air flotation device by the addition of between about 10 parts per million and about 75 parts per million on a dry weight basis of acrylamide/Ethanaminium, N,N,N-trimethyl-2-((1-oxo-2-propenyl)oxo)-, chloride copolymer, said acrylamide/Ethanaminium, N,N,N-trimethyl-2-((1-oxo-2-propenyl)oxo)-, chloride copolymer having a weight average molecular weight ...

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11-01-2018 дата публикации

METHOD FOR FRACTIONATING COMPONENTS OF A BIOMASS OF PROTEIN-RICH MICROALGAE

Номер: US20180007932A1
Автор: Patinier Samuel
Принадлежит: ROQUETTE FRERES

The invention relates to a method for fractionating the components of a biomass of protein-rich microalgae of the genus , characterized in that it comprises the following steps: 1Chlorella,. A method for fractionating components of a biomass of protein-rich microalgae of genus comprising:providing a microalgal biomass produced by fermentation,optionally, washing the biomass so as to eliminate interstitial soluble compounds,thermal permeabilization of the biomass at a temperature of between 50 and 150° C., for a time of between about 10 seconds and 5 minutes,separation between the biomass thus permeabilized and a soluble fraction by a centrifugation technique,optionally, recovery and clarification of the soluble fraction by microfiltration so as to remove residual insoluble substances therefrom, andpurification of the preceding soluble fraction by precipitation, so as to obtain a peptide isolate and a peptide concentrate.2ChlorellaChlorella vulgaris, Chlorella sorokinianaChlorella protothecoides.. The method according to claim 1 , wherein claim 1 , the microalgae of genus are chosen from the group consisting of and3. The method according to claim 1 , wherein the biomass of protein-rich microalgae is prepared by a method which comprises:{'sub': '3', 'a first fermentation step, deficient in nitrogen, in which pH regulation is performed with an NH/KOH mixture, and then'}{'sub': '3', 'a second step of removal of this nitrogen deficiency by a pH regulation performed with NHalone.'}4. The method according to claim 1 , wherein the heat treatment is at a temperature of between about 80 and 150° C. claim 1 , for a time of between about 5 seconds and about 5 minutes.5. The method according to claim 1 , wherein the heat treatment is performed at a temperature of about 85° C. for about 1 minute or at a temperature of about 140° C. for about 10 seconds.6. The method according to claim 1 , wherein the permeabilized biomass is subsequently treated by:milling, preferentially ...

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14-01-2016 дата публикации

CHROMATOGRAPHIC PURIFICATION OF ANTIBODIES FROM CHROMATIN-DEFICIENT CELL CULTURE HARVESTS

Номер: US20160009762A1
Принадлежит:

Methods for the improved purification of antibodies and other proteins from protein preparations including the steps of conditioning the protein preparation by contacting it with multivalent organic ions, then applying the conditioned preparation to an adsorptive chromatography medium. 126.-. (canceled)27. A method for the purification of at least one desired protein from a protein preparation comprising the steps of (a) conditioning an impure protein preparation by contacting it with at least one species of soluble or insoluble multivalent organic ion , subsequently removing solids with the result of removing at least 95% of chromatin , thereby providing a conditioned protein preparation; then (b) applying the conditioned protein preparation , optionally after a buffer equilibration step , to an adsorptive chromatography medium for purification of the desired protein.28. The method of claim 27 , step (a) claim 27 , wherein the conditioning of the protein preparation with at least one species of organic multivalent ion comprises contacting the protein preparation with an electropositive organic additive.29. The method of claim 28 , wherein the electropositive organic additive comprises a soluble electropositive organic additive.30. The method of claim 28 , wherein the electropositive organic additive comprises an electropositive organic cation selected from the group consisting of methylene blue claim 28 , ethacridine claim 28 , chlorhexidine claim 28 , benzalkonium chloride claim 28 , and cetyl trimethyl ammonium bromide.31. The method of claim 28 , wherein the electropositive organic additive is present at a concentration of 0.001% to 2% claim 28 , 0.005% to 1% claim 28 , 0.01% to 0.05% or 0.020% to 0.025%.32. The method of claim 27 , step (a) claim 27 , wherein the conditioning of the protein preparation with at least one species of organic multivalent ions comprises contacting the protein preparation with an insoluble electropositive organic additive in the form ...

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10-01-2019 дата публикации

Method for Processing Solutions of Biomolecules

Номер: US20190009215A1
Автор: Charles HEISE, Tibor Nagy

Apparatus for in-line liquid exchanging a biomolecule-containing liquid is provided. The apparatus comprises a means (3) for mixing at least two liquids comprising a multiple inlet flow-controller (2), the means for mixing also comprising an outlet in fluid connection with a tangential flow filtration device (1) configured in single-pass mode.

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11-01-2018 дата публикации

METHOD FOR PREPARING NANOPRECIPITATES OF LOW MOLECULAR WEIGHT PEPTIDE OR PROTEIN

Номер: US20180009841A1
Принадлежит: Carlina Technologies

The present invention relates to a method for the non-denaturing preparation of peptide or protein nanoprecipitates, or of peptide or protein and metal ion nanocoprecipitales, in which said protein or said peptide has a molecular weight no higher than 20 kDa, preferably no higher than 15 kDa, advantageously no higher than 10 kDa, and more advantageously no higher than 8 kDa. Said method includes a step of preparing a mixture of an aqueous solution of peptides or proteins, a nonsolvent of the peptide or protein, and optionally a water-soluble metal salt. The present invention also relates to a nanoprecipitate that can be obtained by the method according to the invention, as well as to a pharmaceutical composition comprising same, for use in the treatment or prevention of diabetes. 1. Method of non-denaturing preparation of peptide or protein nanoprecipitates or of peptide or protein and metal ion nanocoprecipitates , having a mean diameter of less than 1 μm , comprising the following steps:a) preparation of a mixture of an aqueous solution of peptides or proteins, a nonsolvent of the peptides or proteins, and optionally a water-soluble metal salt;b) gentle stirring of the mixture obtained in step a);c) solid-liquid separation of the mixture obtained in step b); andd) optionally, collection of the peptide or protein nanoprecipitates or the peptide or protein and metal ion nanocoprecipitates,wherein said peptides or proteins have a molecular weight of 20 kDa or less, preferably of kDa or less, advantageously of 10 kDa or less, more advantageously of 8 kDa or less,and said nonsolvent is selected from polyethylene glycols or polyethylene glycol derivatives having a molecular weight of less than 2,000 Da, advantageously between 200 and 2,000 Da, more advantageously of 550 Da, and organic diols selected from the group of hexylene glycol, butane-1,4-diol, pentane-1,5-diol, ethohexadiol, 2-methylpentane-2,4-diol(hexylene glycol), 3-cyclopentene-1,2-diol, cis-4-cyclopentene-1 ...

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11-01-2018 дата публикации

Arginine-rich Peptide Mixture, their Application Thereof in Cervical Cancer Therapy, and a Process for Producing same

Номер: US20180009842A1
Принадлежит:

A process for producing an arginine-rich peptide mixture and the application thereof in cervical cancer therapy is provided. The process includes the following steps: A suspension of walnut meal and egg albumin is pretreated with ultrahigh pressure, and then digested by alkaline proteinase and papain in separated steps with the ultrasonic and microwave-assisted extraction. The peptides of interest are isolated from filtration supernatant obtained after the enzyme digestion by reversed phase high-performance liquid chromatography. By using the peptide mixture as a template, acrylic acid and methyl acrylic acid as functional monomers, triethylene glycol dimethacrylate as cross-linking agent, and isopropylthioxanthone in acetone as a photoinitiator, polymerization is induced by ultraviolet light to form a surface imprinted membrane for isolating and enriching the peptides of interest from the supernatant. The arginine content in the peptide mixture is more than 18%. The arginine-rich peptide mixture is able to strongly suppress the proliferation of human cervical cancer Hela cells. The approach is applicable to reduce the cost of production and speed up the commercialization of large-scale production. 1. A process for producing an arginine-rich peptide mixture , comprising the following steps:(a) collecting a supernatant comprising peptides of interest, wherein de-fatted and pulverized walnut meal, egg albumin and water are mixed and stirred, pretreated with ultrahigh pressure, and subsequently enzymatic digested with ultrasonic-microwave-assisted extraction, wherein enzymes are inactivated by raising temperature and the supernatant is collected through plate and frame pressure filtration; and(b) isolating the peptides of interest from the supernatant, wherein the supernatant is freeze-dried, and the peptides of interest in freeze-dried coarse powder are subsequently isolated by using reversed phase high-performance liquid chromatography (RP-HPLC), wherein Everest C18 ...

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11-01-2018 дата публикации

MODULATION OF CHARGE VARIANTS IN A MONOCLONAL ANTIBODY COMPOSITION

Номер: US20180009876A1
Принадлежит:

Combinations of different chromatography modalities with particularly refined conditions significantly reduce acid charge variants in a preparation of monoclonal antibodies. The process for reducing acid charge variants utilizes a combination of anion exchange and hydrophobic interaction chromatography, followed by cation exchange chromatography polishing, whereby the levels of acidic or basic charge species of the monoclonal antibodies may be modulated to a desired level. 1. A process for removing acid charge variants from a monoclonal antibody , comprising(a) loading a mammalian cell-expressed monoclonal antibody preparation onto a Protein A support, and eluting the monoclonal antibody from the Protein A support, thereby producing a first eluate comprising the monoclonal antibody;(b) loading the first eluate from step (a) onto an anion exchange and hydrophobic interaction (AEX/HIC) chromatography support, and allowing the first eluate to flow through the support, thereby producing a flow-through pool comprising the monoclonal antibody;(c) loading the flow-through pool comprising the monoclonal antibody onto a cation exchange (CEX) chromatography support having an antibody binding capacity of from about 25 g/L to about 65 g/L, determining when the absorbance units measured at UV A280 decrease from about 7% to about 14% from the peak absorbance units measured at UV A280, and then washing the CEX chromatography support with a wash buffer having a pH of from about 5.8 to about 6.6 and a conductivity target of from about 6.6 mS/cm to about 7.6 mS/cm; and(d) eluting the monoclonal antibody from the CEX chromatography support in step with an elution buffer having a pH of from about 6.0 to about 6.4 and a conductivity target of from about 10 mS/cm to about 14 mS/cm, thereby producing a second eluate comprising the monoclonal antibody and from about 10% to about 20% by weight of acid charge variants of the monoclonal antibody.2. The method of claim 1 , wherein the ...

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14-01-2021 дата публикации

FULL FLOW-THROUGH PROCESS FOR PURIFYING RECOMBINANT PROTEINS

Номер: US20210009634A1
Принадлежит:

The present invention concerns a method for purifying a protein comprising in a continuous mode: one filtration step involving the use of at least one chelating agent, an exchanging step involving the use of at least one diafiltration membrane, and a polishing step involving the use of a combination of membrane adsorbers, wherein two membrane adsorbers of said combination are orthogonal in terms of mechanism of action. 2. The method of claim 1 , wherein the filtered protein solution obtained at the end of step (a) is directly passed over the at least one diafiltration membrane claim 1 , without undergoing any treatment such as pH adjustment claim 1 , buffer exchange or dilution.3. The method of claim 1 , wherein the retentate obtained at the end of step (b) is directly passed over said combination of membrane adsorbers claim 1 , without undergoing any treatment such as pH adjustment claim 1 , buffer exchange or dilution.4. The method of claim 1 , wherein the method does not comprise any intermediate storage between the three steps (a) claim 1 , (b) and (c).5. The method of claim 1 , to wherein the method has a flowpath functionally closed.6. The method of claim 1 , wherein only one buffer is used over the whole purification method.7. The method of claim 1 , wherein the one buffer comprises Tris claim 1 , Tris-HCl claim 1 , Bis Tris claim 1 , phosphate and/or citric acid claim 1 , in particular comprises or consists of (i) Bis Tris claim 1 , Tris or Tris-HCl claim 1 , (ii) acetic acid claim 1 , (iii) water and (iv) optionally salt.8. The method of claim 1 , wherein the at least one chelating agent matrix of the filtration step is selected from the group consisting of activated carbons claim 1 , diatomite earth claim 1 , free cationic exchange resin claim 1 , free anionic exchange resin claim 1 , and free mixed mode resin.9. The method of claim 8 , wherein the at least one chelating agent matrix is a combination of two different chelating agent matrices.10. The method ...

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10-01-2019 дата публикации

Polysaccharide Purification for Vaccine Production using Lytic Enzymes, Tangential Flow Filtration, and Multimode Chromatography

Номер: US20190010187A1
Принадлежит: Inventprise, LLC

An improved, cost effective and shortened process of purification of the capsular polysaccharide of , Group B and is disclosed. The process includes a cocktail of enzyme treatment, tangential flow filtration, and multimodal chromatography purification. For Gram-negative bacteria, endotoxin removal process involves endotrap HD resin. This shortened process achieved the purity required by WHO/EP/BP for the use in human vaccine preparation, with simple steps and higher yield as compared to conventional processes. The steps of the process avoid use of organic solvents such as, for example, alcohol, phenol, and ultracentrifugation that are otherwise expensive and time consuming to perform, and/or health hazards for commercial use. This process disclosed is also simple, efficient, non-toxic, easy to scale-up, and environmentally friendly. 1. A process of purifying polysaccharide comprising:providing a fermentation harvest of Gram-positive and/or Gram-negative bacteria;clarifying the fermentation harvest with deoxycholate;concentrating the clarified polysaccharide by a first diafiltration;treating the first diafiltered polysaccharide with an enzyme to digest impurities;precipitating the enzyme with acetic acid;concentrating the polysaccharide by a second diafiltration;passing the second diafiltered polysaccharide through multimodal chromatographic resin and/or endotoxin removal resin; andcollecting the purified polysaccharide.2. The process of claim 1 , wherein the polysaccharide comprises bacterial cell surface polysaccharide.3. The process of claim 2 , wherein the bacterial cell surface polysaccharide comprises capsular polysaccharide and/or exopolysaccharide.4S. pneumoniaeStreptococcus, H. influenzae, S. typhimuriumN. meningitis.. The process of claim 1 , wherein the fermentation harvest comprises claim 1 , Group B and/or5. The process of claim 1 , wherein clarifying is followed by pH adjustment to about pH 3.5-5.0.6. The process of claim 5 , wherein the pH adjustment ...

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10-01-2019 дата публикации

Tangential Flow Filtration Process for Concentrating Biomolecule Solutions

Номер: US20190010189A1
Автор: Heise Charles, Nagy Tibor

A method of concentration of a liquid comprising a biomolecule is provided. The method comprises passing the liquid through a tangential flow filtration device under pressure, wherein the pressure applied is varied between at least a higher pressure and a lower pressure. Preferably, the variation in pressure is delivered by the use of a variable flow-controller, such as a valve. 1. A method of concentration of a liquid comprising a biomolecule which comprises passing the liquid through a tangential flow filtration device under pressure , wherein the pressure applied is cycled between at least a higher pressure and a lower pressure.2. The method according to claim 1 , wherein the method is carried out in-line.3. The method according to claim 1 , wherein pressure is applied by means of a pump claim 1 , and the pressure is varied by operation of a variable flow valve.4. The method according to claim 3 , wherein the variable flow valve is an intermittent flow valve.5. The method according to claim 1 , wherein two different pressures are employed claim 1 , the higher pressure being employed for up to 99.9% of the time of application of the pressure.6. The method according to claim 1 , wherein at least 10 cycles are employed.7. The method according to claim 1 , wherein a cycle frequency of less than 100 Hz is employed.8. The method according to claim 7 , wherein the cycle frequency is from 0.05 to 0.5 Hz.9. The method according to claim 1 , wherein two different pressures are employed claim 1 , and the higher pressure is at least 1.05 fold greater than the lower pressure.10. An apparatus for in-line concentration of a biomolecule-containing liquid comprising a tangential flow filtration device in fluid connection with a means for imparting flow of the liquid through the tangential flow filtration device and a variable flow valve claim 1 , wherein the means for imparting flow is located upstream of the filtration device claim 1 , the variable flow valve is located ...

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10-01-2019 дата публикации

METHOD FOR PURIFYING IMMUNOGLOBULIN SOLUTIONS

Номер: US20190010216A1
Принадлежит:

Herein is reported a method for purifying cell cultivation supernatants either directly after fermentation or after one or more preliminary purification steps, such as protein A affinity chromatography. By adjusting the pH value in the acid range and subsequent incubation of the acidified solution host cell nucleic acid and host cell protein can be precipitated but the target polypeptide remains in solution. Thereafter the precipitate and therewith the contaminating host cell components can be removed by a simple physical separation step. 1. A method for producing an immunoglobulin comprising the following steps:i) adding a solution consisting of an acid and water to a cell cultivation supernatant from which cells and cell debris have been removed for adjusting the pH value to a value of from pH 4.5 to pH 5.5, whereby the solution is essentially free of divalent cations,ii) incubating the pH adjusted cell cultivation supernatant, andiii) removing the precipitate from the incubated cell cultivation supernatant and thereby producing the immunoglobulin,wherein the cell cultivation supernatant comprises the immunoglobulin at a concentration of not more than 10 mg/ml,wherein the concentration of the added acid is 5.5 mol/l or lower.2. The method according to claim 1 , wherein at least 90% of the immunoglobulin remains in solution during the incubating step.3. The method according to claim 1 , wherein the incubating is at a temperature of from 2° C. to 10° C.4. The method according to claim 3 , wherein the incubating of the pH adjusted cell cultivation supernatant is at a temperature of about 4° C.5. The method according to claim 1 , wherein the incubating the pH adjusted supernatant is for about at least 2 hours.6. The method according to claim 5 , wherein the incubating is for about 2 hours to about 72 hours.7. The method according to claim 6 , wherein the incubating is for about 2 hours to about 48 hours.8. The method according to claim 7 , wherein the incubating is ...

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14-01-2021 дата публикации

METHOD OF PREPARING CORN GLYCOPEPTIDES, AND PRODUCT AND USE THEREOF

Номер: US20210010048A1
Принадлежит: QIQIHAR UNIVERSITY

A method for preparing corn glycopeptides, and a product and use thereof, and belongs to the technical field of food and medicine production. The method for preparing corn glycopeptides includes the preparation of corn peptides powder and the glycosylation reaction between corn peptides and amino sugar; the preparation raw material of the corn peptides powder is a corn protein rich in branched chain amino acids; and the corn peptides and the amino sugar are subjected to the glycosylation reaction catalyzed by a transglutaminase, and finally the corn glycopeptides is obtained. The method for preparing corn glycopeptides provided by the present disclosure is simple; and the prepared corn glycopeptides have antagonistic and protective effects on alcoholic liver injury, and can be applied to the preparation of related foods, medicines and health care products. 1. A method for preparing corn glycopeptides , comprising the following steps:(1) hydrolyzing a corn protein suspension, conducting liquid-solid separation, and drying a liquid phase to obtain corn peptide powder;(2) mixing the corn peptides powder with water to formulate a corn peptides solution with a mass-volume concentration of 2-15%;(3) adding amino sugar into the corn peptides solution to obtain a reaction base solution; wherein the addition amount of the amino sugar is 2-5 times of the mass of the corn peptides powder;(4) adding a transglutaminase into the reaction base solution to enable a glycosylation reaction between the corn peptides and the amino sugar, so as to obtain the corn glycopeptides after the reaction is finished; wherein the addition amount of the transglutaminase is 40-80 U of the transglutaminase per 1 g of the corn peptides powder.2. The preparation method according to claim 1 , wherein the hydrolytic enzyme in the step (1) comprises one or more of a Alcalase protease claim 1 , a Protamex protease and a Flavourzyme protease claim 1 , the hydrolysis temperature is 55-65° C. claim 1 , the ...

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09-01-2020 дата публикации

PROTEIN PRODUCTION METHOD

Номер: US20200010503A1
Принадлежит:

Provided herein are methods of producing a heterologous polypeptide and compositions comprising same. 1. A method of producing a heterologous polypeptide , the method comprising the steps of:a) growing Gram-negative bacteria comprising a nucleotide sequence encoding a heterologous polypeptide operably linked to an inducible promoter under fed-batch fermentation conditions in a synthetic medium;b) inducing expression of the heterologous polypeptide;c) harvesting the bacteria in the synthetic medium by decanting;d) homogenizing the bacteria contained in the synthetic medium;e) obtaining from the synthetic medium comprising the homogenized bacteria of step d) a soluble fraction comprising the heterologous polypeptide and an insoluble fraction comprising the heterologous polypeptide;f) resuspending the insoluble fraction;g) centrifuging the resuspended insoluble fraction of step g) to obtain a supernatant comprising the heterologous polypeptide;h) denaturing the polypeptide in the supernatant obtained from step g);i) refolding the denatured polypeptide of step h); andj) subjecting the refolded polypeptide to anion exchange chromatography to obtain the heterologous polypeptide.2. The method of claim 1 , further comprising the steps of:k) precipitating the heterologous polypeptide from the soluble fraction of step e) in a precipitate;l) resuspending the polypeptide precipitate from step k);m) centrifuging the resuspended polypeptide precipitate to obtain a supernatant comprising the heterologous polypeptide;n) subjecting the supernatant obtained in step m) to tangential flow filtration to form a filtration product;o) performing a two-phase separation on the filtration product of step n) to produce an aqueous polypeptide phase and a detergent phase;p) denaturing the polypeptide in the aqueous polypeptide phase obtained in step o);q) refolding the denatured polypeptide of step p); andr) subjecting the refolded polypeptide to anion exchange chromatography to obtain the ...

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03-02-2022 дата публикации

METHODS FOR PRODUCING AFLIBERCEPT IN CHEMICALLY DEFINED MEDIA HAVING REDUCED AFLIBERCEPT VARIANTS

Номер: US20220033471A1
Принадлежит:

The present disclosure pertains to compositions comprising anti-VEGF proteins and methods for producing such compositions. 1. A polypeptide having an isolated amino acid sequence comprising at least 70% sequence identity over a full length of said isolated amino acid sequence as set forth in the group consisting of SEQ ID NO.: 2 , SEQ ID NO.: 3 , SEQ ID NO.: 4 , SEQ ID NO.: 5 , SEQ ID NO.: 6 , SEQ ID NO.: 7 , SEQ ID NO.: 8 , SEQ ID NO.: 9 , SEQ ID NO.: 10 , SEQ ID NO.: 11 , SEQ ID NO.: 12 , SEQ ID NO.: 13 , SEQ ID NO.: 14 , SEQ ID NO.: 15 and SEQ ID NO.: 16 , wherein said polypeptide excludes IdeS as set forth in SEQ ID NO.: 1.2. The polypeptide of claim 1 , wherein said isolated amino acid sequence has at least 80% sequence identity over a full length of said isolated amino acid sequence as set forth in the group consisting of SEQ ID NO.: 2 claim 1 , SEQ ID NO.: 3 claim 1 , SEQ ID NO.: 4 claim 1 , SEQ ID NO.: 5 claim 1 , SEQ ID NO.: 6 claim 1 , SEQ ID NO.: 7 claim 1 , SEQ ID NO.: 8 claim 1 , SEQ ID NO.: 9 claim 1 , SEQ ID NO.: 10 claim 1 , SEQ ID NO.: 11 claim 1 , SEQ ID NO.: 12 claim 1 , SEQ ID NO.: 13 claim 1 , SEQ ID NO.: 14 claim 1 , SEQ ID NO.: 15 and SEQ ID NO.: 16 claim 1 , wherein said polypeptide excludes IdeS as set forth in SEQ ID NO.: 1.3. The polypeptide of claim 1 , wherein said isolated amino acid sequence has at least 90% sequence identity over a full length of said isolated amino acid sequence as set forth in the group consisting of SEQ ID NO.: 2 claim 1 , SEQ ID NO.: 3 claim 1 , SEQ ID NO.: 4 claim 1 , SEQ ID NO.: 5 claim 1 , SEQ ID NO.: 6 claim 1 , SEQ ID NO.: 7 claim 1 , SEQ ID NO.: 8 claim 1 , SEQ ID NO.: 9 claim 1 , SEQ ID NO.: 10 claim 1 , SEQ ID NO.: 11 claim 1 , SEQ ID NO.: 12 claim 1 , SEQ ID NO.: 13 claim 1 , SEQ ID NO.: 14 claim 1 , SEQ ID NO.: 15 and SEQ ID NO.: 16 claim 1 , wherein said polypeptide excludes IdeS as set forth in SEQ ID NO.: 1.4. The polypeptide of claim 1 , wherein said polypeptide is capable of cleaving a target ...

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03-02-2022 дата публикации

Anti-vegf protein compositions and methods for producing the same

Номер: US20220033472A1
Принадлежит: Regeneron Pharmaceuticals Inc

The present disclosure pertains compositions comprising anti-VEGF proteins and methods for producing such compositions.

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19-01-2017 дата публикации

METHOD FOR PURIFYING IMMUNOGLOBULIN

Номер: US20170015732A1
Принадлежит: GREEN CROSS HOLDINGS CORPORATION

The present invention relates to a method for purifying an immunoglobulin, and more particularly, to a method for purifying an immunoglobulin, which comprises: dissolving immunoglobulin-containing plasma protein fraction I+II+III or fraction II+III; adding caprylate to the solution to cause precipitation; performing dialysis and concentration after removal of the precipitate; performing anion exchange resin and ceramic cation exchange resin purification processes to effectively remove a solvent and detergent added to inactivate viruses; and performing elution while maintaining salt concentration at a constant level to maintain the immunoglobulin polymer content at a low level. According to the method for preparing the intravenous immunoglobulin according to the present invention, a precipitation step of preparing fraction II from fraction I+II+III or fraction II+III as a starting material can be omitted, and problems, including a complicated process and a low yield, which occur in the conventional preparation process employing the polyethylene glycol treatment process, can be solved by use of first sodium caprylate precipitation, anion exchange chromatography and cation exchange chromatography. In addition, when the immunoglobulin purification method according to the present invention is used, the efficiency with which impurities and thrombotic substances are removed can be increased and the immunoglobulin polymer content can be maintained, and thus a stable immunoglobulin with increased quality can be produced. 1. A method for purifying an immunoglobulin , comprising the steps of:(a) dissolving immunoglobulin-containing plasma protein fraction I+II+III or fraction II+III, followed by performing a precipitation reaction by adding a precipitant;(b) removing a precipitate produced from (a), followed by filtering a supernatant comprising immunoglobulin, concentrating a filtrate, subjecting a concentrate to an anion exchange chromatography, and recovering a fraction not ...

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21-01-2016 дата публикации

Production of pulse protein product

Номер: US20160016991A1
Принадлежит: Burcon Nutrascience MB Corp

A pulse protein product having a protein content of at least about 50 wt % (N×6.25) d.b. is recovered in the processing of pulse protein source material to form pulse protein products wherein the pulse protein source is extracted in one embodiment with calcium salt solution. The resulting pulse protein solution is separated from the bulk of the residual pulse protein source and then the pulse protein solution is processed to remove finer residual solids, which are optionally washed and then dried to provide the pulse protein product. In another embodiment, the pulse protein source is extracted with water, the bulk of the residual protein source removed and the resulting pulse protein solution treated with calcium salt to precipitate phytic acid. The precipitated phytic acid and any finer residual solids remaining in solution after the initial separation step are removed from the pulse protein solution then optionally washed and dried to provide the pulse protein product.

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