METHOD FOR PROVIDING INFORMATION FOR DIAGNOSING NEURODEGENERATIVE DISORDER

30-08-2018 дата публикации
Номер:
KR1020180096981A
Принадлежит:
Контакты:
Номер заявки: 00-17-102023470
Дата заявки: 22-02-2017

[1]

The present invention refers to method for the diagnosis of a neurodegenerative disorders selected from ball number information, method more particularly diagnosis of Alzheimer's disease, a diagnostic kit and diagnostic device and, in particular nerve [eyk cow petty extracted through analysis of diagnosis of Alzheimer's disease Amyloid beta are disclosed.

[2]

[3]

Alzheimer's disease is effected while truncated Amyloid precursor protein (amyloid precursor protein) which causes inclusion of Alzheimer's disease Amyloid beta (amyloid a-β, A β) and is stored, is accumulated A β generate active oxygen (ROS, reactive oxygen species) of mitochondria activate mitosis, this activates the action of Alzheimer's disease to induce BACE1 (beta-a amyloid converting enzyme 1) protein, BACE1 owing to the activation of action is provided to the neuronal cell damage and, pharmaceutical compositions containing as the loss of a language ability to uniformly brain fingerprinting causing diseases, as well as brain fingerprinting disorders, preferably death serious territory make known.

[4]

In order existing diagnosis of Alzheimer's disease primarily during A β Amyloid beta1-42, A β or1 - 40 Measures the amount of deflection, said ELISA method was used in order to measure the volume of material. In the case of ELISA substrate can hold N terminal (or C - terminal) placing the acquisition antibodies, for measuring the concentration of detection antibody after binding the C - terminal (or N - terminal) Amyloid beta where it is, in the case of Alzheimer's disease related thought that the Amyloid beta multimers, C - container to reference terminal subsequently, if existing method to diagnose Alzheimer's disease, multimers detection antibody reaction of Amyloid beta and C - terminal are reacted door number becomes very small as the substrate. This is outside the displayed Amyloid beta is multimers can react the C - terminal happened reduces are disclosed. The, chamber number the same concentration of the corresponding detection signal even if a value measured according to the amount of Amyloid beta multimers to one of advantageous use the concentration of Amyloid beta variation detection.

[5]

In order to solve said C - terminal door number dependent on reaction with the progressing direction of light but without using antibodies, ELISA or part of same index measuring electrochemical sensor disconnecting decides antibodies display converter, using C - terminal are not necessarily required in the nanometer range. In addition, without using C - terminal N - terminal judged A β that in some electrochemical sensor1- X Which is reacted with, existing plasma (plasma) using the same detection in the diagnosis of Alzheimer's disease has been incorrect number point at the door that result.

[6]

[7]

1. 한국등록특허 제100595494호

[8]

The present invention refers to different neurodegenerative disorders including Alzheimer's disease to diagnose number for information are disclosed.

[9]

The present invention refers to Alzheimer's disease and other neurodegenerative disorders or predictive prognosis or diagnosis, or a novel method, biomarkers, kit and device object number also are disclosed.

[10]

[11]

One embodiment form of the present invention, a subject (i) containing the biological sample to yield a vesicle (vesicles); (ii) using antibodies specific for the Amyloid beta of N - terminal, measuring the level of Amyloid beta included in said biological sample; and (iii) for comparing the measured level of said levels showed sample prepared; including a, information number ball method for the diagnosis of a neurodegenerative disorders are disclosed.

[12]

The present invention refers to said biological sample of Amyloid beta (iv) compared with control samples said regulated by detecting increased levels, confirming whether said subject is neurodegenerative disorders; can be further comprises.

[13]

The present invention refers to said biological sample obtained from a vesicle (vesicle) identical or different; further including a predetermined time interval lapses disclosed.

[14]

Said [eyk cow petty (exosome) is a vesicle (vesicle), fine particles, fine vesicles, nano some (nanosome), extracellular vesicles and (ectosome) least one selected from the group consisting of process from.

[15]

Said nerve - derived [eyk cow petty, astrocytes - derived [eyk cow petty, oligodendrocyte - derived cells selected from the group consisting of glue and fine [eyk cow petty - derived [eyk cow petty process from least one.

[16]

(Neuronal exosome) derived from said nerve - [eyk cow petty preferably disclosed.

[17]

Antibodies that bind said N - terminal specific, Amyloid beta N - be a combination with the terminal.

[18]

Antibodies that bind said N - terminal specific, Amyloid beta 1 to 16 times or some bio-sequence listing for engaging with the N - terminal side is more than once.

[19]

The level of Amyloid beta said, Amyloid beta protein, protein phosphorylation, the game be miRNA or mRNA levels.

[20]

Said biological samples are blood, plasma, serum, saliva and urine process from selected from the group consisting of least one.

[21]

Said neurodegenerative disorders Alzheimer's disease (AD), vascular dementia, dementia (FTD) fronts, cortical base regression (CBD), progress nuclear as a matter of paralysis (PSP), dementia with Lewy element, knot - dominant senile dementia (tangle-a predominant senile dementia), (Pick ' sdisease) (PiD), hu granules (argyrophilic grain) diseases, amyotrophic lateral sclerosis (ALS) Huntington, other movement disorders, interested (Guam)- treating Parkinson dementia composite, FTDP provided 17, cell - body lysine (Lytico provided Bodig disease), multiple sclerosis, traumatic (TBI) and Parkinson's disease selected from the group consisting of process from.

[22]

Preferably said neurodegenerative disorders (AD) in Alzheimer's disease.

[23]

Process from fixed connected to said biosensor antibodies.

[24]

Said EIS a biosensor includes a sensor array (Electrochemical impedance spectrometry), bead based EIS sensor, ELISA (enzyme linked immunoassay), SPR (Surface Plasmon Resonance) biofeedback-based assay sensor and FET (Field a-effect transistor)Sensor selected from the group consisting of be a production cost.

[25]

Measuring the level of said biological samples contained in the Amyloid beta, N - terminal of an antibody specific for Amyloid beta Amyloid beta react with biological samples contained in said after, impedance (impedance) change or current (current) is more than for measuring changes.

[26]

40% To 80% ratio of impedance change said back, said subject is confirming that has neurodegenerative disorders; can be a.

[27]

Current ratio of 1 to 20% said back, said subject is confirming that has neurodegenerative disorders; including a is more than.

[28]

[29]

Other embodiment forms of the present invention, diagnosis or prognosis in a subject or neurodegenerative disorders, identifying or a subject at risk of neurodegenerative disorders, neurodegenerative disorders in a subject or treating benefit from therapy with prescribing as predicting method, a vesicle (vesicles) (i) containing a subject biological sample to yield a; (ii) using antibodies specific for the Amyloid beta of N - terminal, measuring the level of Amyloid beta included in said biological sample; and (iii) for comparing the measured level of said levels showed sample prepared; including a, method are disclosed.

[30]

[31]

Another embodiment of the present invention form, subject as a set biomarkers for diagnosing neurodegenerative disorders, subject biological samples containing said vesicle (vesicles) included for measuring the level of Amyloid beta, including antibodies specific for Amyloid beta of said N - terminal characterized biomarker set are disclosed.

[32]

[33]

Another embodiment of the present invention forms, as a kit for diagnosing neurodegenerative disorders subject, subject biological samples containing said vesicle (vesicles) for measuring the level of Amyloid beta included, said kit including antibodies specific for Amyloid beta of N - terminal characterized are disclosed.

[34]

[35]

Another embodiment form of the present invention, a subject biological sample containing a vesicle (vesicles) (i) is disposed between the stator inlet; (ii) Amyloid beta specific N - terminal and including antibodies, said antibodies reactive with a biological sample is introduced into said inlet portion; and (iii) the mode for reactions in said reaction, said Amyloid beta of a measuring biological samples contained features; a diagnosis device including neurodegenerative disorders are disclosed.

[36]

[37]

The present invention refers to different neurodegenerative disorders including Alzheimer's disease can be number information for diagnosis.

[38]

The present invention refers to Alzheimer's disease and other neurodegenerative disorders or predictive prognosis or diagnosis, or a novel method, biomarkers, kit and device also can be number.

[39]

[40]

Figure 1 shows a bead filler or nanoparticle surface antibodies to an, an electrochemical sensor for measuring the amount of Amyloid beta [eyk cow petty spinal cord derived from abstract indicating method are disclosed. Figure 2 shows a sensor surface antibodies also resistance/current based electrochemical method for measuring the amount of Amyloid beta [eyk cow petty fixed spinal cord derived from a general outline are disclosed. Figure 3 shows a 1 to one or more of beads disposed within the microwell traps also patterned microwell indicating that general outline are disclosed. Figure 4 patterned microwell with chamber number representing the antibodies are disclosed. Figure 5 shows a bead by reacting antibodies present in Amyloid beta also, indicating the general outline for measuring impedance that are disclosed. Figure 6 plasma (plasma), and neural [eyk cow petty [eyk cow petty derived from antibodies in order to identify an element is processed to distinguish Amyloid beta of bead surface represents the value measuring device using the result in a change in impedance of are disclosed. Figure 7 shows a change in impedance in a sample of two (left panel) and ROC 47 also determining the inherent sensitivity and specificity (right panel) indicating the result value measuring chart are disclosed. Figure 8 shows a fixed reduced (reduced graphene oxide) [eyk cow petty GO also antibodies binding to Amyloid beta peptides derived from surface indicating the general outline method are disclosed. Figure 9 shows a human plasma (plasma) is sampled in a subject sample also decomposition of extracting Amyloid beta antibodies are fixed in extracting reduced GO reacting to measuring the amount of Amyloid beta abstract for indicating method are disclosed. Figure 10 reduced GO sensor determining the result indicating resistance are disclosed.

[41]

Specific method described herein, protocol, cell lines, analysis and reagent can be changed since, not limited them to understand that the present invention refers to accomplishing. The specific embodiment of the present invention terms as used herein for describing aspects of the present invention and not for limiting the range of a appended claims and not won't understand accomplishing.

[42]

As used in the specification and appended claims, otherwise not specify context, indicating that a single may be accomplishing recognizing that an object comprising a plurality. The, for example, of a plurality of such fragments and referred "fragments", "antibody" includes one or more antibodies and must therefore referred to publicly known splicing in of this equalized referred of are disclosed.

[43]

Not otherwise defined, all terms and scientific terms as used herein in the present invention relative to conventional techniques with the flawless typically by the same semantics and it will have the meanings. Method and materials described herein can be used in any method of the present invention embodiment or testing and material similar or equivalent but, in a preferred method, device and material is hereinafter disclosed. The present invention disclosure document are associated with all citation herein used in document disclosure reported method, reagent and means for purposes of disclosure herein described on the right side as a whole introduced into the substrate. The present invention is eligible to anticipating this preceding invention none herein through this disclosure be interpreted as admit that no don't substrate.

[44]

Alternatively displayed not, techniques of the present invention embodiment is therefore detect future, biochemical, molecular biology, cellular biology, mutating, immunology and pharmaceutical conventional use a method are disclosed.

[45]

[46]

The present invention refers to a vesicle (vesicles) [eyk cow petty biomarker or biomarker partially analyzed, for example, Alzheimer's disease (AD), dementia (FTD) neurodegenerative disorders including multiple sclerosis (MS) and fronts may have a subject which is likely to generate a identified by the discovery that are disclosed.

[47]

The present invention refers to neurodegenerative diseases (e.g., Alzheimer's disease) partially present in the circulatory system of a subject having non-neural - derived [eyk cow petty certain biomarkers (e.g., Amyloid beta) discovery is not expected of increased based on substrate. The present invention refers to the biomarkers of neurodegenerative disease in a subject by analyzing a level [eyk cow petty diagnosing neurodegenerative disorders can be demonstrated as follows.

[48]

The present invention refers to certain biomarkers in a subject using measurement of neurodegenerative diseases derived from neural - [eyk cow petty to subsequent occurrence prediction (e.g., neurodegenerative disorders danger of generating identifying a subject) can be also's desire.

[49]

The present invention measures the amount of Amyloid beta tested in a sample of a patient undergoing a Alzheimer's disease victims of the automatically change, depending on the type of sample between said normal and that a patient has been changed of dividing. I.e., to treat patients with normal plasma (plasma) when sample when [eyk the cow petty is defined between distinct than have, particularly nerve [eyk the cow petty is down after confirming the possible than would clear, the present invention the arrears of work (reference 6 also). I.e., as samples for measuring the amount of Amyloid beta than plasma, from the surface of the sample (or neural [eyk cow petty) [eyk cow petty as contained therein to the patient as a normal amount of Amyloid beta distinguish displeased.

[50]

[Eyk cow petty it with patient because of Amyloid beta according to the amount of normal system capable of, the present invention refers to measuring the amount of Amyloid beta [eyk cow petty it with, said measured in comparison with a predetermined reference amount by reference (reference), distinguish between patient and normal flow tides.

[51]

Extracting said be a in a biological sample. And, biological sample said blood, plasma, serum, saliva and urine can be a selected from the group consisting of cost, the one number are not disclosed. In addition, more preferably in said nerve [eyk cow petty (neural exosome). In this case, measuring the amount of Amyloid beta in, between distinct to treat patients with normal further alluding to him.

[52]

The present invention refers to subject's neurodegenerative evaluating the condition biomarkers set number substrate. The embodiment embodiment, biomarker levels are analyzed substrate. The present invention refers to the amount of Amyloid beta characterized [eyk cow petty it with measurement, measuring the amount of Amyloid beta said 802.11a packets one method are specially number but not, generated after reaction with antibodies, a change in impedance or current for measuring changes is more than (also 6 and 7 also reference).

[53]

In one embodiment of the present invention in, said impedance change is 20% to 90%, or 30% to 80%, or 40% to 80%, or 40% to 70%, or 50% to 60%, or 55% to 70% the game back and Alzheimer's disease, preferably 60 to 65% to prevent the back can be Alzheimer's disease. In addition, 0 said current change. 1% To 20%, or 0. 1% To 10%, or 1% to 9%, or 2% to 8%, or 3% to 7%, or 4% to 6% the game back and Alzheimer's disease, preferably 3 to 5% the game back be Alzheimer's disease. The present invention incorporated in the embodiment and carry to the victims of the experiment, this numerical range therefrom.

[54]

The present invention refers to compositions described herein can be used in method number substrate. These compositions comprise small molecule compounds; antibody or a functional active fragments including peptides and proteins; and small interfering ribonucleic acid (siRNA), micro - RNA (miRNA), ribozyme and anti - sense sequences comprising including can be polynucleotides (for example, document [Zeng (2003) ProcNatl Acad Sci USA 100:9779 - 9784]; and document [Kurreck (2003) Eur J Biochem 270:1628 - 1644] reference).

[55]

The present invention refers to diagnosis or prognosis or neurodegenerative disorders in a subject, of identifying or a subject at risk of neurodegenerative disorders, neurodegenerative disorders in a subject having or treating benefits from prescribing therapy methods and kits for predicting the number substrate. The embodiment in aspects, of one or more antibodies specifically bind [eyk cow petty kit, a biomarker of one or more antibodies specifically bind, collecting biological samples/for holding one or more vessels, and kits comprising the signs for use.

[56]

The aim organized the neck is only for short number and as used herein, any scheme described herein taking into way defining don't substrate.

[57]

[58]

Biological samples

[59]

The present invention refers to Alzheimer's disease and other neurodegenerative disorders biomarkers for diagnosis and prognosis method and a number substrate. Biomarker levels can be measured in biological samples are obtained from a subject. Some embodiment in aspects, of the present invention can be obtained from biological sample is blood. Some embodiment in aspects, about 1 to 10 ml of blood sampled from a subject substrate. In another embodiment embodiment, about 10 to 50 ml of blood sampled from a subject substrate. Blood is arm, leg, or central venous catheter can be accessed through including blood, can be sampled from any suitable region of the body.

[60]

Some embodiment in aspects, treating or activity after blood collection with each other. For example, blood is medical can be sampled after. Sampling time so as to increase the number and/or compositions used to also can be adjusted. For example, inducing blood vascular expansion can be cut to exercise or therapy.

[61]

Subsequent techniques to prepare or preserving blood sample after the taking of various components can be combined with. For example, some embodiment in aspects, blood is after the taking of anticoagulant number, cells fixed number, number number number comprised of billion, billion nucleotide sequence number number number phosphate, protein, DNA or RNA preservation number treated substrate. Some embodiment in aspects, blood is anticoagulant number, for example, EDTA or heparin by using vacuum collection tube containing intravenous puncture through collection with each other. Blood heparin - coated and subcutaneous injection syringe needle sampled by disapproval. Blood is cell culture may be useful in combination with may be disclosed. For example, some embodiment in aspects, blood is cell culture media or cell culture media supplemented with (e.g., cytokines) in combination with each other.

[62]

Biological samples are whole blood, serum, plasma, urine, interstitial fluid, peritoneal fluid, cervical swab specimen, tears, saliva, [hyep side swab specimen, skin, cerebral spinal fluid, or for example, other brain tissue including tissue including, in publicly known source must therefore obtained from other disapproval.

[63]

[64]

Vesicles ([Eyk cow petty, Fine particles, fine vesicles, Some nano, Extracellular Vesicles and ) Concentrates or isolates

[65]

Sample positive selection, voice selection, negative or positive selection via an audio selection can be to concentrate vesicles. Some embodiment in aspects, the parcel directly capture with each other. In another embodiment embodiment, blood cell vents and, biological samples are collected from the remaining the parcel. Some embodiment in aspects, in a biological sample the concentrated the parcel [eyk cow petty, fine particles, fine vesicles, nano some, extracellular vesicles or among others. Some embodiment in aspects, a lipid vesicle comprising a concentrated in biological samples derived from neural - [eyk cow petty, astrocytes - derived [eyk cow petty, derived oligodendrocyte - [eyk cow petty and fine glue cells [eyk cow petty derived - among others.

[66]

The basis of the differences in biochemical properties sample is vesicles to concentrate vesicles may be disclosed. For example, sample antigen, nucleic acid, metabolism, the basis of the differences in gene expression or epigenesis (epigenetic) to concentrate vesicles can be. Antigen difference based on aspects in some embodiment, flow measuring cells with antibodies of bonded - field gradient in ferrimagnetic or paramagnetic bead or fluorescence labelled antibodies are used. Nucleic acid difference based on aspects in some embodiment, flow measuring cells are used.

[67]

Based on metabolic difference aspects in some embodiment, flow cells or another classification number times measured by dye intake/measuring techniques are used. In some embodiment aspects based on gene expression, cell culture using cytokines are used. In other publicly known biochemical properties must therefore sample to concentrate based on vesicles may be disclosed. For example, sample is based on pH or mobility to concentrate vesicles can be. Further, some embodiment in aspects, this parcel concentrated more than one method are used. In another embodiment embodiment, sample is an antibody, ligand or soluble receptor by using vesicles are to concentrate.

[68]

In another embodiment embodiment, the sample surface markers used in the concentrated positive vesicles are used. Some embodiment in aspects, the parcel [eyk cow petty, fine particles, fine vesicles, nano some, extracellular vesicles or among others. In another embodiment embodiment, NCAM, CD171, CD9, CD63, CD81, various nerve or astrocytes attached proteins, fine glue cells CD18/11, or CD3 T cell surface markers used concentrated cell membranes are used. Some embodiment in aspects, vesicle population not found in cell surface markers on cells by vesicles used has been concentrating depleting are used. Measuring extracellular or intracellular or cell surface marker fluorescence-labeled joined to flow cells also classification using the marker further concentrating can be used. Intracellular marker and preferentially expressed in intracellular or extracellular markers vesicles extracellular proteins can be nuclear dye or antibody. [Eyk cow petty preferentially expressed on the cell surface markers (e.g., NCAM) antibodies to cell surface antigen can be. Some embodiment in aspects, for example cell surface markers, including nerve - CD171 NCAM or a derived [eyk cow petty surface markers are disclosed. Some embodiment in aspects, monoclonal NCAM, CD9, CD63, CD81 or CD171 concentrated or isolating antibodies from a sample used are used. In a particular embodiment, NCAM, CD9, CD63, CD81 or CD171 antibodies encoded bio neel mote. The embodiment embodiment, the streptavidin - agarose resin or beads or CD171 antibodies by using bio it became neel mote NCAM subsequently can be isolated antibodies [eyk cow petty - can be capable of forming a complex. In another embodiment embodiment, NCAM, CD9, CD63, CD81 CD171 antibodies or monoclonal anti - human NCAM, CD9, CD63, CD81 or CD171 antibodies are disclosed.

[69]

Some embodiment in aspects, a lipid vesicle comprising a concentrated desiccant from the biological samples for the specific type of vesicles subsequently concentrated substrate. For example, biological samples are to concentrate and [eyk cow petty, then concentrated nerve - [eyk cow petty derived for subsequently concentrated substrate. Some embodiment in aspects, biological samples are vesicles to concentrate source individual neuronal cells are disclosed. In a particular embodiment, vesicles neuronal source is fine glue cells, nerve or astrocytes are disclosed.

[70]

In another embodiment embodiment, the vesicle binding to biological samples the parcel under conditions which present the parcel number number - number number capable of forming a complex biological samples on said contacting said number number; and said vesicle - number number from the composites said vesicles isolated said vesicles isolated from a biological sample including method to yield a sample containing or concentrated and, the biological samples said purity vesicles used to purity greater than present vesicles.

[71]

Embodiment specific embodiment, an antibody or a lectin is said number number are disclosed. Vesicle - lectin composite is a dielectric ceramic disclosed patent application disclosure number 2012/0077263 useful intermediates in this call. Some embodiment in aspects, the parcel [eyk cow petty, fine particles, fine vesicles, nano some, extracellular vesicles or among others. Some embodiment in aspects, nerve - derived [eyk cow petty, astrocytes - derived [eyk cow petty, derived oligodendrocyte - [eyk cow petty - [eyk cow petty or fine glue cells derived from among others. Some embodiment in aspects, a plurality of isolated or concentrated steps are carried out. The embodiment then in a particular embodiment, isolating step performed isolated from blood sample number 1, number 2 isolated from other [eyk cow petty step carried out by isolating neural - derived . In another embodiment embodiment, vesicle - number number composite for one time portion is dissolved by reagent soluble, dissolved vesicles protein levels are analyzed substrate. Some embodiment in aspects, antibodies generated on heparin - vesicle complex. In another embodiment embodiment, said method further comprising releasing the package from the antifoaming composite - antibodies. Specific embodiment embodiment, the solid non - magnetic beads, magnetic beads, agarose or cephalosporin loss are disclosed. In another embodiment embodiment, the parcel antibody - vesicle complex 3. 5 To 1. 5 Exposing the low pH is released by other. In another embodiment embodiment, high pH solution released by adding a lipid vesicle comprising a neutralized substrate. In another embodiment embodiment, emitted vesicles with nocodazole emitted the parcel dissolved solution dissolved with each other.

[72]

In another embodiment embodiment, dissolved solution is comprised of salt forms of number and comprising nucleotide sequence number for number number billion.

[73]

[74]

Neurodegenerative disorders

[75]

The present invention refers to diagnosis or prognosis or neurodegenerative disorders in a subject, of identifying or a subject at risk of neurodegenerative disorders, neurodegenerative disorders in a subject or treating benefit from therapy with predicting method for prescribing a number substrate.

[76]

Some embodiment in aspects, neurodegenerative disorders Alzheimer's disease (AD), vascular dementia, dementia (FTD) fronts, cortical base regression (CBD), progress nuclear as a matter of paralysis (PSP), dementia with Lewy element, knot - dominant senile dementia, (PiD), hu granules diseases, proximal sclerosis (ALS), other movement disorders, such as Parkinson's interested - dementia composite, FTDP provided 17, cell - lysine body, multiple sclerosis, traumatic brain injury (TBI) and Parkinson's disease selected from the group consisting of brain damage.

[77]

Some embodiment in aspects, the present invention refers to one or more neurodegenerative disorders diagnosis or prognosis in a subject enables doctors to the print disclosed. In another embodiment embodiment, the present invention refers to one or more neurodegenerative diseases or as a stand-alone diagnostic possibility enables doctors to number times the number can be. In another embodiment embodiment, the present invention refers to generate enables doctors to neurodegenerative disorders can be identify risk subject. In another embodiment embodiment, the present invention refers to generating enables doctors knowing what the subject is neurodegenerative disorders can be brings. Additional embodiment embodiment, the present invention refers to enables doctors to neurodegenerative disorders in a subject having or treating benefits from the prescribing therapy can be predicted.

[78]

[79]

Biomarkers

[80]

(For example, Alzheimer's disease) or biomarker level neurodegenerative disorders have biological samples obtained from a subject having a risk for developing in an other. Some embodiment in aspects, Amyloid beta biomarkers are disclosed. N - terminal can be as biomarkers of the present invention refers to among all Amyloid beta. The specification (including drawing) Amyloid beta antibodies specific for N - terminal is in "Abeta 1 a-x" prevent the big Abeta. 1 - 16 Of the forefoot of Amyloid beta (Abeta) at arbitrary scan line can be N - terminal acting as both. E.g. Abeta intermediate 3 3 from N - terminal acting as a memory area for urging the front even once is pivotably. N terminal is reached once severed after 6 - 39 6 acting, once C - terminal 39 can be act.

[81]

In another embodiment embodiment, Tau for development, 1 Aβ provided 42, TDP-a 43, α - carried out by the inverter, SOD-a 1, FUS, FKBP51, IRS provided 1, IRS-a 1 for development, CTSD, LAMP1, UBP, HSP70, NSE, NFL, CD9, CD63, CD81 and CD171 are disclosed.

[82]

Some embodiment in aspects, of the present invention expression gene of a biomarker of biomarker levels can be measured by other. Specific embodiment embodiment, gene expression changes are measured by measuring the expression level of the Amyloid beta. In a particular embodiment, of a biomarker of gene expression is PCR, microarray or sequencing using the anti-glare layer substrate. Some embodiment in aspects, the expression level of the mRNA of a biomarker of the biomarkers are measured by determining levels or miRNA.

[83]

In must therefore is under illumination controller of the present invention can be used as a detecting marker of and analyzed by various method and device knowing those disclosed. Patient test sample of polypeptides or mort, immunoassay device and method are often used substrate. The device and method are various sandwich, competitive or non - competitive assay format using labeled molecules in the presence or quantity of analyte of interest can be associated with that generates a signal. Further, the specific method and device, e.g., the presence or quantity of labeled molecules without the need for a biosensor and optical immunoassay is measuring analyte can be used.

[84]

Other method (for example, measurement of marker RNA levels) are splicing in the user pulls but must therefore is publicly known, preferably by using the biomarker is an immunoassay analysis with each other. The presence or quantity of marker using antibodies that are specific for each marker generally and are identified by detecting specific binding. Any suitable immunoassay, for example, enzyme immunoassay (ELISA) - connected, radiation immunoassay (RIA), competitive binding analysis, such as planar waveguide (planar waveguide) techniques can be used. Antibodies specific immunological binding is directed to a marker can be detected directly or indirectly. Antibodies direct markers attached to fluorescence or luminescence tag, metal, dye, etc. radionuclides. Indirect markers must therefore publicly known well in various enzymes, for example, alkaline phosphate nucleotide sequence number, etc. it ladles and the rock it is sour hose d [swi number.

[85]

The present invention also taken into account by the use of antibodies that are specific for markers fixed with each other. Antibodies solid support various conductors, for example, magnetic or chromatography matrix particles, the surface of the place (e.g., microtiter well) analysis, such as the solid substrate material (e.g., plastic, nylon, paper) pieces can be fixed on. Analyzing strip is an antibody or a plurality of antibodies on the solid support array by coating the number bath 1308. Then, the strip is impregnated into test sample after washing and detection via a measurable signal rapidly processed so as to, for example, colored spot (spot) can be generate.

[86]

A plurality of markers can be separately or simultaneously perform analysis of the test sample. For efficient processing for multiple samples of the various markers can be combined into a test. In addition, splicing in the same user must therefore from multiple individuals (for example, in successive time instants) is worth testing will recognize a plurality of samples. This alternation of change in test series of samples over time will be identifying marker levels. Increased or decreased level of the marker as well as the approximate time from start event (event) identification of portions of changes level marker, the presence and quantity of tissue structure (salvageable), the validity of drug treatment, effect of various therapeutic compounds, an event of identifying severe degrees, severe degrees identifying diseases, and future event including but not limited to them as a result of validating a patient risk of, useful information for disease States will number.

[87]

In the present invention and combinations thereof analysis referred markers associated with different diagnostic through the relevant number can be . The two such panel 1, two 2, two 3, 4 two, two 5, two 6, 7 two, two 8, 9 two, two 10, 15 20 by using one or two or more separate markers can be established. Single marker, or greater than the present invention including a method to form a subset of markers in the marker analysis is carried out in a variety of clinical situations clinical sensitivity or specificity can be optimizing. P-a S312 provided IRS-a 1 and using (R or insulin resistance index) of P-a panY provided IRS provided 1 neurodegenerative diseases can be predicting the risk of or diagnosis.

[88]

Marker also analysis of various physical formats can be performed. For example, microtiter plates or automated using the large number of test samples can be hereinafter for processing. Alternatively, a single sample format through the Internet for example, in a timely manner foreign transport thread or emergency room for immediate treatment and diagnosis to hereinafter can be. In particular physical format useful for a plurality of different analyte detection of a plurality of discrete addressable locations comprises surfaces (addressable). This format is more protein microarrays, or "protein chip" and capillary device without using a tool.

[89]

(For example, Alzheimer's disease) of the present invention biomarkers neurodegenerative disorders in plays an important role in the early detection and monitoring a plurality of hierarchies. Such disorders typically found in a sample can be measured marker body material are disclosed. And a metered quantity is inferior vena cava base disorder or disease pathologies, the presence or absence of neurodegenerative disorders, neurodegenerative disorders may have future probability of correlation. Status of patients undergoing treatment in a metered quantity is correlation are disclosed have transient response to a treatment.

[90]

Some embodiment in aspects, biomarkers immunohistochemistry, immunocytochemical, immune fluorescent, immune precipitation, measured by ELISA method selected from the group consisting of the [wey the [su it shook off and with each other.

[91]

[92]

Clinical analysis performance

[93]

Of the present invention method provides a method for diagnosing or prognosis and/or neural degenerative disorders, neurodegenerative disorders and/or identifying a subject at risk of, neurodegenerative disorders in a subject or treating with profile for predicting clinical benefit from therapy can be used in assays. The sensitivity of the analysis performance clinical analysis, specificity, ROC curve (AUC), accuracy, positive predicted value (PPV) and voice (NPV) can be evaluated by measuring predicted values. Neurodegenerative disorders in a subject or diagnosis or prognosis, identifying or a subject at risk of neurodegenerative disorders, neurodegenerative disorders in a subject or treating benefit from therapy with profile for predicting the analysis herein disclosure in the nanometer range.

[94]

Based on said analyzing clinical performance is sensitive can be. The sensitivity of analysis of the present invention at least about 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100% implementation being. Based on said analyzing clinical performance also relates can be. The assay of the present invention at least about 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100% implementation being. ROC curve (AUC) can be based on said analyzing clinical performance. AUC assay of the present invention is at least about 0. 5, 0. 55, 0. 6, 0. 65, 0. 7, 0. 75, 0. 8, 0. 85, 0. 9 Or 0. Wednesday 95 1. Said analysis clinical performance accuracy can be based on disclosed. The washing of the present invention can include at least about 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100% implementation being.

[95]

[96]

Composition

[97]

Useful in the composition of the present invention method specifically recognize biomarkers associated with neurodegenerative disorders comprising composition, said Amyloid beta biomarkers are disclosed.

[98]

Some embodiment in aspects, said composition comprises at least one Amyloid beta activity of the base. In another embodiment embodiment, said composition comprises at least one Amyloid beta active from the process chamber. In another embodiment embodiment, said composition peptide, nucleic acid, antibodies and small molecule selected from the group consisting of.

[99]

Specific embodiment embodiment, the present invention refers to a composition specifically detected biomarkers associated with neurodegenerative disorders are disclosed.

[100]

Some embodiment in aspects, composition comprising antibodies, antibodies specifically bind to said biomarkers or vesicles of the present invention. The present invention refers to as biomarkers using Amyloid beta, among N - terminal characterized as biomarkers of Amyloid beta are disclosed. To this end, said antibodies specific for Amyloid beta N - terminal preferably disclosed. I.e., Amyloid beta with the terminal N - said antibodies can be disclosed.

[101]

As used herein the term "antibody" further discussed hereinafter and biomarkers or vesicle (for example, [eyk cow petty) reacts specifically the event is for containing a fragment thereof are disclosed. Antibodies using techniques can be fragmented by conventional, fragment thereof is the same as manner for a whole lie lower screening 1308. For example, F (ab) 2 fragment thereof is antibodies can be produced by treatment with pepsin. The resulting F (ab) 2 fragment thereof is reduced to produce compound Fab fragments can be crosslinked. Recombinant DNA technique also antigen-binding portions thereof, or can be produced by enzymatic or chemical cleavage of intact antibodies. The antigen-binding portions thereof in particular Fab, Fab ', F (ab') 2, Fv, dAb and complementarity determining regions (CDR) fragments, single chain antibodies (scFv), single domain antibodies, bispecific antibodies, chimeric antibodies, humanized antibodies, diaryl body, and specific antigen binding polypeptides so as to impart sufficient immunoglobulin polypeptides containing at least a portion of having a predetermined wavelength. Specific embodiment embodiment, antibodies can be detected and attached to an indicator (for example, the cover is radioisotope, fluorescent compound, enzyme or enzyme cofactor viscoelastic) further comprises a substrate.

[102]

Specific embodiment embodiment, of the present invention antibodies and monoclonal antibodies, in one aspect the present invention refers to of the present invention specifically bind to novel biomarkers specific embodiment [eyk cow petty or generating a method available substrate. For example, monoclonal antibodies that specifically bind the biomarker or biomarker or [eyk cow petty [eyk cow petty generating method, or fragments thereof including immunogenic composition, in an amount effective to stimulate an immune response detectable administering mouse, antibody-producing cells (for example, from spleen cells) obtained from a mouse antibody-producing cells with myeloma cells to yield a fusion antibody species other than said generating, and said biomarkers or [eyk cow petty generating antibodies capable of specifically binding to a species other than testing a hybridoma producing the monoclonal antibodies can be confirming comprising. Once the obtained, optionally high - [eyk cow petty monoclonal antibodies specifically bind biomarkers derived from hybridoma cells or proliferating in culture conditions generating cell cultures can be. Monoclonal antibody can be used from cell culture can be positive number.

[103]

In antibodies referred to generally at understanding terms used in "specific specifically reactive" must therefore as antibodies of interest (for example, bio-markers or [eyk cow petty) and between other antigen antigen without interest for sufficiently selective means that are disclosed. Methods for using the specific method, for example, in therapeutic applications, can be preferably higher than the degree of specificity in binding. Generally desired antigen monoclonal antibody (monoclonal antibodies as compared to multiple) greater than effectively identifying cross-reacting polypeptides have a tendency. Antibody: antigen specificity of the interaction affecting properties on the antibodies to an antigen affinity are disclosed. A variety of different affinity desired specificity can be reached but, generally preferably about 10 antibodies-6 , 10-7 , 10-8 10 Or-9 Hereinafter have affinity (dissociation constant) are disclosed.

[104]

For example antibodies, derived from neural - [eyk cow petty, including Amyloid beta, [eyk cow petty or biomarkers of the present invention specifically bind epitope of can be generated.

[105]

Further, in a preferred antibodies in order to identify the technique involves screening antibodies used to affect the nature of the antibodies can be obtained. Interaction between a variety of different techniques in particular antibodies antigen prepared by using identifying preferred antibodies can be used. These techniques ELISA, surface plasmon resonance binding assay (for example, via core (Biacore) binding assay, via core Abbe (Biacore AB) (Sweden [ep it burns, material)), sandwich analysis (for example, child mouse n international (IGEN International, Inc.) of paramagnetic beads (American compared to [su bug material main gating) system), the [wey the [su it shook off blot, immune precipitation analysis, immunocytochemical and comprises immunohistochemistry.

[106]

Some embodiment in aspects, the present invention refers to compositions used for treating or preventing neurodegenerative disorders are disclosed.

[107]

Detailed as described elsewhere herein, the present invention refers to a wide variety of neurodegenerative disorders are Amyloid beta, for example, Alzheimer's disease is associated with the discovery that pathologies based on substrate.

[108]

[109]

Treatment method

[110]

The present invention refers to an effective amount of composition including the step of administering to a subject, said method as neurodegenerative disorders in a subject, said composition the level of Amyloid beta, a method number substrate. In another embodiment embodiment, composition comprises at least one Amyloid beta activity of the base. In another embodiment embodiment, returning composition comprises at least one Amyloid beta activity. In another embodiment embodiment, composition peptide, nucleic acid, antibodies and small molecule selected from the group consisting of.

[111]

[112]

Kit

[113]

Another aspect of the present invention a kit for detecting or monitoring the P19 in a subject neurodegenerative disorders. Various kits are the present invention are taken into account by different components. Generally speaking, said means of at least one biomarker in a subject kit are provided to will. In another embodiment embodiment, the biological sample kit said collecting means, means of at least one biomarker in a biological sample are provided to, and kit for use contents of the signs will be disclosed. Specific embodiment embodiment, said means for isolating or concentrating kit in the biological sample. In further embodiments, concentrated or isolated from a biological sample or concentrated isolating means comprising reagents needed. In a particular embodiment, said kit includes means for quantifying the amount of biomarkers. In further embodiments, means for determining the volume of a biomarker of reagents required for detecting the quantity of a biomarker of 2000.

[114]

[115]

Hereinafter, the present invention through further detailed in the embodiment to less than 1000. The present invention more specifically account for these in the embodiment is provided, in the embodiment of the present invention range and not the limited to these.

[116]

[117]

In the embodiment 1. [Eyk cow petty Preparing samples

[118]

1. 1. In blood [Eyk cow petty(Exosome) Extraction

[119]

In the blood to extract, a number ratio of 1:3 and reagent mixing and re-deposition onto a stand-alone first blood plasma calcium, the thrombin solution every other number of retard depletion method of plasma calcium re-deposition onto said reacted with a reagent is mixed. The precipitating solution is obtained from steam centrifuging, the supernatant (supernatant) was PBS dilution to retard is number for reparing over. In order [eyk cow petty supernatant dilution ExoQuick separating said dilution only with a solutionTM Solution by mixing 1 with respect to the reaction time. Then, the pellets in a stand-alone number stored in the supernatant (pellet) centrifuge are obtained. As well as keeping them in the container whereupon the extracted PBS solution.

[120]

[121]

1. 2. Nerve [Eyk cow petty(Neural Exosome) Extraction

[122]

Said 1. 1. Since derived from extracted from various cells, nerve [eyk cow petty positive number of abortion process in order difference 2 are extracted. Said 1. 1. 3% BSA in PBS solution (Bovine Serum Albumin) have dilution is prepare [eyk cow petty solution is blended, a dilution solution (Biotinylated) nerve cell membranes treated biotin specific protein [eyk cow petty reacted reaction antibody solution. Nerve cell membranes which are selectively used for gripping specific protein reaction antibodies are disclosed. Then, 40 μl of streptavidin - agarose resin (Streptavidin Agarose Resin) 20 μl of 3% BSA solution made into solution, 4 °C in 10 minutes with respect to the reaction. The reacted solution is centrifuging the supernatant number been stand-alone. Biotin - streptavidin - agarose resin for separating glycine - in hydrochloric acid buffer (Glycine a-HCl buffer solution, 50 mm, pH 3. 0) Added to reduce pH, centrifuging is carried out at room temperature (Vortexer) spray enough behind his line. Present new container and transferred to an supernatant, - tris hydrochloric acid buffer (Tris a-HCl buffer solution, 1M, pH 8. 6) Stored in the neutral by the addition of a solution. Said positive number nerves in PBS solution by a process [eyk cow petty solution is 80 °C whereupon after completion as well as experiments.

[123]

[124]

1. 3. Nerve [Eyk cow petty inA β 1-A x Extracting method

[125]

In the case of modified danger of Amyloid beta (A β) is connected as well as by extracting immediately before measuring. First, melting [eyk cow petty solution is about 10 minutes 4 °C 80 °C nerve being stored value, adding 1:1 RIPA buffer solution at a rate of 4 °C [eyk cow petty-melt while maintaining his 1 minutes sonication. [Eyk cow petty film are degraded by the process while Aβ 1 a-x said mixed solution is extracted to gain.

[126]

[127]

In the embodiment 2. Bead A method of treating spinal cord or nanoparticle surface antibodies A β extracted from cow petty1 A-x Detection

[128]

2. 1. Beads Small number on which an upper arm

[129]

The present invention small number for victims of the bead using sensor, first, i) carrying a layer of silicone dioxide oxide etching silicon substrate shape patterned electrode by using light, ii) an electron beam deposition using titanium - raising the number remaining portion is a stand-alone been platinum electrode to lift-off process. Iii) then, electrode separation utilizing Su8 vulcanization-based photoresist patterning for future beads Microwell [wan line structure with a lower sensor and, iv) about 2 mm thickness for forming a PDMS well for introducing external sample PDMS aromatic vinyl to 2 - 3 mm of holes have been produced. V) PDMS well device including the bonding oxygen to plasma treatment process is performed, and form a lower sensor bonding PDMS well finished his. Figure 3 shows a microwell traps located within at least one of beads 1 to patterned microwell also enables general outline degrees and, representing the chamber number 4 are disclosed.

[130]

[131]

2. 2. Method

[132]

Antibodies surface reacts with a functional group capable of activated micro beads (M provided 280 Tosylactivated magnetic beads, Dynabeads yarn) solution (200 μl) at ambient temperature after lamination centrifugal separated beads, beads magnets immobilize and the supernatant was a stand-alone number. 0. 0 1 Mm phosphate buffer solution. 2 Μm diameter and with every other porous membrane filter, the remaining beads was added 1 ml of buffer. Said process wash process to eliminate other residues as beads are disclosed. 3 Times after repeated washing of supernatant and number and a stand-alone, 720 μl of 0. 80 Μl of antibody solution added was 1 mm phosphate solution sufficiently intermingled. In 24 hours 37 °C beads solution is obtained by reacting said process, two types of using a buffer after bead solution to wash-gate. Said two types of buffer of the like filter prepared PBS buffer and Tris buffer and the line, a stand-alone number after supernatant prepared bead solution, PBS buffer at room temperature by adding 5 minutes of electrons have intermingled well, 37 °C 4 in time with respect to the latter Tris using a buffer to wash after reaction. Wash-stand-alone number again the supernatant solution bead fluorescence, 200 μl of PBS solution was filtered into the remaining beads 4 °C of the installed application.

[133]

Bead solution is PBS buffer solution at a rate of preparation of 1:150 dilution have, in the embodiment 1 that have been extracted from said bead solution at a rate of 40 minutes 1 Aβ provided x prepare 1:2 with respect to the reaction solution. Reaction beads magnets immobilize and the supernatant after number been stand-alone. Then, PBS buffer solution by washing the usage have intermingled well, only PBS solution 3 was prepared by washing once more complete bead before measuring.

[134]

For bead solution is dropped into the element, respectively patterned microwell to be located beneath a patterned microwell shaft beads in the briefcase. Then, an active light emitting electrochemical measuring change in impedance (Autolab PGSTAT302N) were measured.

[135]

[136]

2. 3. Result

[137]

In order to identify and use utility [eyk cow petty, A β plasma (plasma), and neural [eyk cow petty [eyk cow petty by extracting for A β [eyk cow petty and nerve [eyk cow petty capable of dividing database identifying whether derived from him. In the case of an impedance change is extracted from plasma (plasma) A β differs regulated not formate, derived from patients with Alzheimer's disease [eyk cow petty A β can be ascertained in the case of impedance change has been the extra. Placed between the first and, in the case of nerve [eyk cow petty A β derived from patients with Alzheimer's disease compared to [eyk cow petty A β derived from rapidly year without impedance change has been confirmed that the difference to the second unambiguously assigned in overlapping section (6 also).

[138]

The present invention is to make sure that the victims of the results of the number of samples of said clearly than two character is increased to said 47 such as a long time. As a result, result of said impedance change in Alzheimer's disease (AD) patient (NC) and similarly regulated by a discrete (left panel of Figure 7) has been confirmed. This extremely high precision as compared to existing blood based method in that it exhibits, using 100% and 90% sensitivity and degree of specificity for ROC chart in each measured as signal peptides (right panel of Figure 7).

[139]

The, nerve [eyk cow petty antibodies with bead or nanoparticles in extracting Amyloid beta by measuring a change in impedance so produced and offers high accuracy of diagnosis of Alzheimer's disease has been confirmed that the patient can be connected.

[140]

[141]

In the embodiment 3. Resistance/current based electrochemical sensor - FET and resistive sensor

[142]

3. 1. Reduced GO (reduced Graphene Oxide, RGO) Sensor used for the preparation of a number - rGO thin film coating and sensor

[143]

(Graphene oxide) is ultrasonic decomposition method according GO Modified Hummers method number is small fine piece plate has been peeled off. The strip has a relatively large centrifugal separation to anger, using only the relatively thinner plate has a transparent conductive layer, Milli a-Q water was stored in deionized water. The MDD GO thin film is silicon oxide coated substrate carrying a layer of silicone dioxide (Meniscus-a dragging Deposition) method. First, preparing glass have no residue formed directly processing buzzed antagonist activity, prepare the silicon substrate and the substrate solution to sizable GO glass terminals in said solution, 30 degrees angle while maintaining a constant speed (20 mm/s) was nickel. Said process can be repeated one or more times when the 20, said thin film made by gaseous hydrogen iodide in acid 3 applied to a GO GO 80 °C reduction in time rGO obtained thin film. Then, the remaining photoresist mask making rGO using thin film optical etching process using oxygen plasma reactive ion or acrylic acid number was a stand-alone (300 W, 30 sec). After etching the remaining mask number was a stand-alone lift-off process, the remaining rGO thin film optical etching process for connecting both ends again - alloys have been produced gold electrode pattern. Said electrodes raise away line and the portion remaining after the electron beam deposition. A thin film made rGO element stored in the padded back functionality with respect to the activated oxygen plasma rGO antibodies.

[144]

[145]

3. 2. Reduced GO (reduced Graphene Oxide, RGO) Sensor used for the preparation of a - surface treatment

[146]

20 Μl rGO made for thin-film portion of EDC (1 a-ethyl-a 3 - [3 a-dimethylaminopropyl] - carbodiimide, 2 mm, Sigma Aldrich, USA) solution, NHS (N-a succinimide, 8 mm, Sigma Aldrich, USA) solution, 10 mm PBS solution with respect to the exposed during time of 1. After antibody solution (1 Aβ provided x 100,102,104,106 antibodies, antibody 6E 10, Abcam yarn) and EDC, NHS rGO reacting antibodies is configured to receive time using a mixed solution 2 to the surface.

[147]

[148]

3. 3. Method

[149]

In nerve [eyk cow petty sample extracted from 1 Aβ provided x have prepared in the embodiment 1, in the embodiment 3. 1 And 3. 2 RGO sensor is provided so as to prepare the in, 1 Aβ provided x 30 minutes with respect to the prepared sample to the reaction surface. Then, Semiconductor parameter analyzer equipment resistance were measured.

[150]

[151]

3. 4. Result

[152]

30 Sample of extracting neural , nerve [eyk cow petty A β extracted from using his database. As a result, a change in the resistance (resistance change) in Alzheimer's disease (AD) patient compared to high percentage of regulated has been confirmed that the lower electrode.

[153]

The, in extracting Amyloid beta antibodies are fixed by reacting rGO nerve [eyk cow petty sensor by measuring resistance change has been confirmed that the possible diagnosis of Alzheimer's disease.



[1]

The present invention relates to a method for providing information for diagnosing a neurodegenerative disorder. The method comprises the steps of: (i) obtaining a biological sample containing vesicles from a subject; (ii) measuring a level of amyloid beta contained in the biological sample using an antibody specific for the N-terminal of amyloid beta; and (iii) comparing the measured level to a level of a control sample which has been prepared. The present invention can provide information for diagnosing neurodegenerative disorders including Alzheimer′s disease.

[2]

COPYRIGHT KIPO 2018

[3]

[4]

  • (AA) Antibody (N-terminal)
  • (BB) A β _1-x derived from nerve exosomes (or exosomes)
  • (CC) Beads or nanoparticles
  • (DD) Fix the antibody to the surface of the beads (or nanoparticles)
  • (EE) Amyloid β reaction derived from the nerve exosomes (or exosomes)
  • (FF) Results
  • (G1,G2) Electrochemical sensor
  • (HH) Locate the beads on the electrochemical sensor
  • (II) Measurement



(I) containing a subject biological sample to yield a vesicle (vesicles); (ii) using antibodies specific for the Amyloid beta of N - terminal, measuring the level of Amyloid beta included in said biological sample; and (iii) for comparing the measured level of said levels showed sample prepared; including a, information number ball method for the diagnosis of neurodegenerative disorders.

According to Claim 1, said biological sample of Amyloid beta (iv) compared with control samples said regulated by detecting increased levels, confirming whether said subject is neurodegenerative disorders; including a characterized the, ball method number information for the diagnosis of a neurodegenerative disorders.

According to Claim 1, said biological sample obtained from a vesicle (vesicle) identical or different; characterized in further including the, ball method number information for the diagnosis of a neurodegenerative disorders.

According to Claim 3, said [eyk cow petty (exosome) is a vesicle (vesicle), fine particles, fine vesicles, nano some (nanosome), extracellular vesicles and (ectosome) least one selected from the group consisting characterized, information number ball method for the diagnosis of neurodegenerative disorders.

According to Claim 4, said nerve - derived [eyk cow petty, astrocytes - derived [eyk cow petty, derived oligodendrocyte - [eyk cow petty and fine glue - least one selected from the group consisting of cells derived [eyk cow petty characterized, information number ball method for the diagnosis of neurodegenerative disorders.

According to Claim 4, characterized in that said neural - derived [eyk cow petty (neuronal exosome), information number ball method for the diagnosis of neurodegenerative disorders.

According to Claim 1, said N - terminal specific antibodies that bind, Amyloid beta characterized by combination with the N - terminal, information number ball method for the diagnosis of neurodegenerative disorders.

According to Claim 1, said N - terminal specific antibodies that bind, Amyloid beta binding part or entire bio-sequence listing N - terminal side 1 to 16 times once characterized, information number ball method for the diagnosis of neurodegenerative disorders.

According to Claim 1, said level of Amyloid beta, Amyloid beta protein, protein phosphorylation, characterized in that the mRNA or miRNA level, information number ball method for the diagnosis of neurodegenerative disorders.

According to Claim 1, said biological samples are blood, plasma, serum, saliva and urine management devices selected from the group consisting of least one characterized, information number ball method for the diagnosis of neurodegenerative disorders.

According to Claim 1, said neurodegenerative disorders Alzheimer's disease (AD), vascular dementia, dementia (FTD) fronts, cortical base regression (CBD), progress nuclear as a matter of paralysis (PSP), dementia with Lewy element, knot - dominant senile dementia (tangle-a predominant senile dementia), (Pick ' sdisease) (PiD), hu granules (argyrophilic grain) diseases, amyotrophic lateral sclerosis (ALS) Huntington, other movement disorders, interested (Guam)- treating Parkinson dementia composite, FTDP provided 17, cell - body lysine (Lytico provided Bodig disease), multiple sclerosis, traumatic (TBI) and Parkinson's disease selected from the group consisting characterized, information number ball method for the diagnosis of neurodegenerative disorders.

According to Claim 11, characterized in that said neurodegenerative disorders Alzheimer's disease (AD), information number ball method for the diagnosis of neurodegenerative disorders.

According to Claim 1, characterized in that said antibody is fixed connected to biosensor, information number ball method for the diagnosis of neurodegenerative disorders.

According to Claim 13, said EIS a biosensor includes a sensor array (Electrochemical impedance spectrometry), bead based EIS sensor, ELISA (enzyme linked immunoassay), SPR (Surface Plasmon Resonance) biofeedback-based assay sensor and FET (Field a-effect transistor)Characterized sensor selected from the group consisting of cost, information number ball method for the diagnosis of a neurodegenerative disorders.

According to Claim 1, measuring the level of said biological samples contained in the Amyloid beta, N - terminal of an antibody specific for Amyloid beta Amyloid beta react with biological samples contained in said after, impedance (impedance) change or current (current) measuring change characterized, information number ball method for the diagnosis of neurodegenerative disorders.

According to Claim 15, said impedance change ratio of 40% to 80% back, said subject is confirming that has neurodegenerative disorders; including a characterized the, ball method number information for the diagnosis of a neurodegenerative disorders.

According to Claim 15, 1 to 20% current ratio of said back, said subject is confirming that has neurodegenerative disorders; including a characterized the, ball method number information for the diagnosis of a neurodegenerative disorders.

Neurodegenerative disorders in a subject or diagnosis or prognosis, identifying or a subject at risk of neurodegenerative disorders, neurodegenerative disorders in a subject or treating benefit from therapy with prescribing as predicting method, a vesicle (vesicles) (i) containing a subject biological sample to yield; (ii) using antibodies specific for the Amyloid beta of N - terminal, measuring the level of Amyloid beta included in said biological sample; and (iii) for comparing the measured level of said levels showed sample prepared; including a, method.

Subject biomarkers for diagnosing neurodegenerative disorders as a set, said subject biological samples containing a vesicle (vesicles) included for measuring the level of Amyloid beta, including antibodies specific for Amyloid beta of said N - terminal characterized biomarker set.

As a kit for diagnosing neurodegenerative disorders subject, subject containing said vesicle (vesicles) for measuring the level of Amyloid beta contained in the biological sample, said kit including antibodies specific for Amyloid beta of N - terminal characterized.

(I) containing a vesicle (vesicles) subject biological sample inlet inlet; (ii) Amyloid beta specific N - terminal and including antibodies, said antibodies reactive with a biological sample is introduced into said inlet portion; and (iii) the mode for reactions in said reactor, said Amyloid beta included features of a measuring biological samples; including neurodegenerative disorders a diagnosis device.



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