KR101873971B1 - Diagnostic composition of alzheimer's disease using neuregulin-1 protein and diagnostics method of alzheimer's disease using the same - Google Patents

Abstract

The present invention relates to a diagnostic composition for Alzheimer using neuregulin-1 protein, in particular, soluble neuregulin-1 protein and a diagnosis method of Alzheimer using the same. More specifically, neurengulin-1 is overexpressed in plasma of a patient with Alzheimer and expression amount of the neurengulin-1 increases in proportion to an age of AD patient so that the diagnostic composition for Alzheimer including the neurengulin-1 can be usefully used for efficiently diagnosing Alzheimer.

Description

TECHNICAL FIELD The present invention relates to a composition for diagnosing Alzheimer's disease using neuregulin-1 protein and a method for diagnosing Alzheimer's disease using the same, and a diagnostic method for Alzheimer's disease using the same.

The present invention relates to a composition for diagnosing Alzheimer's disease using a neuregulin-1 protein, specifically a water-soluble neuretin-1 protein, and a method for diagnosing Alzheimer using the same.

With the rapid development of medicine in recent years, the average life span of humans has increased, and as the elderly population has increased, new social problems have been highlighted. In particular, geriatric nervous system diseases such as stroke, Alzheimer's disease (AD), and Parkinson's disease are manifested as dysfunctional nervous system dysfunctions, and so far there is no effective way to treat them.

In particular, AD is the most common neurological disease of the elderly. AD is the most common degenerative brain disease that causes dementia and is a brain disorder that causes memory, thinking and behavioral problems. Dementia is a general term that means loss of memory and other intellectual abilities that are so severe that it interferes with everyday life. AD accounts for 60-80% of cases of dementia.

The pathogenesis of AD and its cause are uncertain, but it is known to be caused by decreased synthesis of the neurotransmitter acetylcholine, deposition of β-amyloid, and damage of nerve cells due to hyperphosphorylation of tau protein. Clinical diagnosis of AD depends mainly on history and neuropsychological tests, and imaging tests such as magnetic resonance imaging (MRI) and PET are performed as secondary tests. PiB-PET (pittaburgh compound E-positron emission tomography) is used to diagnose AD by ascertaining the accumulation level of amyloid beta in the brain through brain imaging, but it is expensive.

In this regard, Korean Patent Laid-Open Publication No. 10-2016-0135430 discloses a method for identifying the expression level of miR-206 in olfactory tissue to diagnose early Alzheimer's disease or mild cognitive impairment.

On the other hand, water-soluble neurengulin-1 (sNRG-1) is a water-soluble form of neurengulin-1 (NRG-1), a member of the neuregulin family and plays an essential role in the nervous system. NRG-1 is synthesized and accumulated in human neocortex, white matter, and brain spinal fluid. While the precursor of NRG is mostly expressed in cortical neurons, sNRG-1 accumulates on the surface of white matter cells.

Biomarkers for early and effective diagnosis of AD should be able to monitor the efficacy of disease interventions during clinical trials and to detect pre-emergence of disease. Therefore, biomarkers based on blood are attracting attention. Recently, several biomarker candidates for the diagnosis of AD have been proposed: a ratio of polymer tau to tau monomer in platelets, a modified pattern of amyloid precursor protein (APP) form, oxidative DNA damage in peripheral leukocytes, plasma 24S- The level of expression of hydroxycholesterol and plasma IL-8, and the like. Although the biomarkers may be biomarkers useful for early AD diagnosis, non-invasively readily available biomarkers with high sensitivity and specificity have not been developed.

Therefore, the present inventors investigated a method for diagnosing Alzheimer's disease using a biomarker usable for diagnosis of AD and the marker, and sNRG-1 protein was overexpressed in Alzheimer's disease patients, which showed a negative correlation with the MMSE score , The MMSE score showed a difference in the expression level in the mild and moderate AD patients, and the expression level was increased as the age of AD patients was increased. Thus, the present invention was completed.

Korean Patent Publication No. 10-2016-0135430

It is an object of the present invention to provide a composition and a kit for diagnosing Alzheimer's disease comprising an agent for measuring the expression level of a neurengulin-1 protein.

It is another object of the present invention to provide a method for providing information on the diagnosis of Alzheimer's disease using the composition for diagnosing Alzheimer's disease.

In order to accomplish the above object, the present invention provides a composition for diagnosing Alzheimer's disease comprising an agent for measuring an expression level of a neurengulin-1 protein.

The present invention also provides a kit for diagnosing Alzheimer's disease comprising the composition for diagnosing Alzheimer's disease.

Furthermore, the present invention relates to a method for measuring the expression level of a neurengulin-1 protein in a sample, And comparing the amount of expression of the neurengulin-1 protein with the amount of neureurin-1 protein expressed in a normal individual. The present invention also provides a method for diagnosing Alzheimer's disease.

The present invention relates to a composition for the diagnosis of Alzheimer's disease. The composition for diagnosing Alzheimer's disease is characterized in that it is overexpressed in plasma of a patient suffering from Alzheimer's disease and the expression amount thereof increases in proportion to the age of AD patients, Can be used effectively.

FIG. 1 is a graph showing Western blot analysis of changes in expression of neuregulin-1, amyloid beta, S100a9, and alpha synuclein protein in plasma of a patient suffering from Alzheimer's disease (CP1: EGF domain of neurengulin-1 protein; CTL: normal sample; AD: Alzheimer's patient sample; CP2: ECD domain of the neuregulin-1 protein).
FIG. 2 is a graph showing the correlation between the amount of neuregulin-1 protein expressed in the plasma of patients with Alzheimer's disease and the MMSE score.
FIG. 3 is a graph showing changes in expression of the neuregulin-1 protein according to age.
4 is a graph showing changes in the expression of the neuregulin-1 protein according to sex.

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for diagnosing Alzheimer's disease comprising an agent for measuring an expression level of a neurengulin-1 protein.

The neurengulin-1 protein is one of the proteins belonging to the neurequiline family, and may be a protein that interacts with the NEU / ERBB2 receptor to increase phosphorylation at the tyrosine residue. The neurengulin-1 protein according to the present invention may be a water-soluble neurengulin-1 protein. As used herein, the term "water-soluble neurengulin-1" refers to BACE1 (beta-secretase cleaving enzyme 1) or gamma-secretase (gamma secretase), which is involved in the production of amyloid- ). ≪ / RTI >

The neurengulin-1 protein may comprise any polypeptide sequence known in the art as the neurengulin-1 protein. The polypeptide may include all variants or fragments of amino acids having different sequences by deletion, insertion, substitution, or a combination of amino acid residues within a range that does not affect the function of the protein. Amino acid exchange in proteins or peptides that do not globally alter the activity of the molecule is well known in the art. And may be modified by phosphorylation, sulfuration, acrylation, saccharification, methylation, farnesylation, or the like depending on circumstances. In one embodiment of the present invention, the neureunin-1 protein may be a polypeptide represented by SEQ ID NO: 1.

The composition that can be included in the composition may be any one or more selected from the group consisting of an antibody, an antibody fragment and an aptamer capable of measuring the expression level of a neurengulin-1 protein. The antibody may be a monoclonal antibody, a polyclonal antibody or a recombinant antibody, which can be easily prepared using techniques known in the art. On the other hand, the antibody fragment may comprise a functional fragment of an antibody molecule. The functional fragment of the antibody molecule means a fragment having at least an antigen-binding function, and examples thereof include Fab, F (ab ') 2, F (ab') 2 and Fv. In addition, the aptamer may be RNA, DNA, modified oligonucleotide or a mixture thereof as an oligonucleotide molecule having a binding activity to a predetermined target molecule. The aptamer may be in a linear or cyclic form.

Such monoclonal antibodies can be prepared using hybridoma methods or phage antibody library techniques known in the art. Generally, hybridoma cells that secrete monoclonal antibodies can be made by fusing immune cells and cancer cells isolated from immunologically appropriate hosts, such as mice injected with antigen proteins. Cell fusion of these two groups can be performed by fusing using methods known in the art, such as polyethylene glycol, and propagating the antibody producing cells by standard culture methods. Specifically, subcloning is performed using a limiting dilution method, and hybridoma cells capable of producing an antigen-specific antibody using the obtained uniform cell population can be prepared by culturing in vitro or in vivo in a large amount. The antibodies prepared by the above methods can be separated and purified by methods such as gel electrophoresis, dialysis, salt precipitation, ion exchange chromatography, affinity chromatography and the like.

The polyclonal antibody can be prepared by injecting an immunogen-causing biomarker protein or a fragment thereof into an external host according to a method known in the art. The external host may be a mammal such as a mouse, rat, sheep, or rabbit. When the immunogen is injected intramuscularly, intraperitoneally or subcutaneously, an adjuvant may be administered together to increase the antigenicity. Thereafter, blood can be routinely taken from an external host to obtain serum showing improved titer and specificity for the antigen, and separating and purifying the antibody therefrom.

The preparation may be a conjugate labeled with a detection agent such as a chromogenic enzyme, a fluorescent substance, a radioactive isotope or a colloid. The chromogenic enzyme may be a peroxidase, an alkaline phosphatase or an acid phosphatase. On the other hand, the fluorescent substance may be at least one selected from the group consisting of fluorescein carboxylic acid (FCA), fluorescein isothiocyanate (FITC), fluorescein thiourea (FTH), 7-acetoxycarin- Yl, 2'7'-dichlorofluorescein-6-yl, dihydrotetramethylresin-4-yl, , Tetramethylrhodamine-5-yl, tetramethylrhodamine-6-yl, 4,4-difluoro-5,7-dimethyl-4-bora-3a, 4a-diaza- -Ethyl or 4,4-difluoro-5,7-diphenyl-4-bora-3a, 4a-diaza-s-indacene-3-ethyl, Cy3, Cy5, polyL-lysine- Isothiocyanate, rhodamine-B-isothiocyanate (RITC), phycoerythrin (PE) or rhodamine.

The agent may further comprise a ligand capable of specifically binding to the agent. The ligand may be a conjugate labeled with a detection element such as a chromogenic enzyme, a fluorescent substance, a radioactive isotope or colloid, and a ligand treated with streptavidin or avidin.

The diagnostic composition of the present invention may contain, in addition to the above-described preparation, distilled water or a buffer solution which stably maintains the structure thereof.

In a specific example of the present invention, the inventors have shown that sNRG-1 protein is overexpressed in Alzheimer's disease patients (see FIG. 1), which has a negative correlation with MMSE scores (see FIG. 2) (See Table 2), and the expression level was increased as the age of AD patients was increased (see FIG. 3). Therefore, a composition comprising the agent capable of measuring the expression level of the NRG-1 protein can be usefully used as a diagnostic composition for Alzheimer's disease.

The present invention also provides a kit for diagnosing Alzheimer's disease comprising the composition for diagnosing Alzheimer's disease.

The composition for diagnosing Alzheimer's disease may have the above-described characteristics. In one example, the composition may comprise an agent that measures the level of expression of the neurengulin-1 protein, which may be any one or more selected from the group consisting of antibodies, antibody fragments, and aptamers. The neurengulin-1 protein may be a water-soluble neurengulin-1 protein, and specifically, the protein may be a polypeptide represented by SEQ ID NO: 1.

The agent may be one that is bound to a solid substrate to facilitate subsequent steps such as washing or separation of the complex. The solid substrate may be made of synthetic resin, nitrocellulose, glass substrate, metal substrate, glass fiber, microspheres or micro beads. In addition, the synthetic resin may include polyester, polyvinyl chloride, polystyrene, polypropylene, PVDF, nylon, and the like.

The kit distinguishes whether a patient is Alzheimer's disease or not, thereby allowing a doctor such as a doctor to diagnose Alzheimer's disease, as well as monitoring the patient's response to the treatment and changing the treatment method according to the result. In addition, an animal model of Alzheimer's disease, including mice and rats, can be used to identify substances that regulate the expression of the neurengulin-1 protein in vitro and in vivo.

The diagnostic kit may be prepared according to a method known in the art, and may further include buffers, stabilizers, inactive proteins and the like as necessary.

In order to search for the amount of the agent for measuring the expression level of the neurengulin-1 protein, the diagnostic kit may be carried out by fluorescence, which is carried out by detecting fluorescence with a fluorescent substance attached thereto as a detection body, or by detecting radiation by attaching radioactive isotopes A high throughput screening (HTS) system using a radiation method, a surface plasmon resonance (SPR) method for measuring the surface plasmon resonance change in real time without labeling the detector, or a SPRI surface plasmon resonance imaging) method.

In a specific example of the present invention, the inventors have shown that sNRG-1 protein is overexpressed in Alzheimer's disease patients (see FIG. 1), which has a negative correlation with MMSE scores (see FIG. 2) (See Table 2), and the expression level was increased as the age of AD patients was increased (see FIG. 3). Therefore, the composition comprising the agent capable of measuring the expression level of the NRG-1 protein can be usefully used for the production of a diagnostic kit for Alzheimer's disease.

Furthermore, the present invention relates to a method for measuring the expression level of a neurengulin-1 protein in a sample, And comparing the amount of expression of the neurengulin-1 protein with the amount of neureurin-1 protein expressed in a normal individual. The present invention also provides a method for diagnosing Alzheimer's disease.

The neurengulin-1 protein may have the characteristics as described above.

The sample can be any biological sample containing a neuregulin-1 protein that can be distinguished from a normal state. Specifically, the sample can be any one selected from the group consisting of urine, blood, serum, and plasma. In one embodiment of the present invention, the sample may be plasma.

The sample can be prepared by a method such as anion exchange chromatography, affinity chromatography, size exclusion chromatography, liquid chromatography, continuous extraction, centrifugation or gel electrophoresis, etc., before measuring the expression level of the neurengulin-1 protein Can be preprocessed.

In the method according to the present invention, the expression level of the neurengulin-1 protein is measured by any one or more methods selected from the group consisting of Western blotting, enzyme-immunochemical detection (ELISA), immunohistochemical staining, immunoprecipitation and immunofluorescence . At this time, if the expression amount of the neurengulin-1 protein is lower than the normal individuals, it can be diagnosed as Alzheimer's disease.

In a specific example of the present invention, the inventors have shown that sNRG-1 protein is overexpressed in Alzheimer's disease patients (see FIG. 1), which has a negative correlation with MMSE scores (see FIG. 2) (See Table 2), and the expression level was increased as the age of AD patients was increased (see FIG. 3). Therefore, the method for measuring the expression level of NRG-1 protein can be useful for providing information for diagnosis of Alzheimer's disease.

Hereinafter, the present invention will be described in detail by the following examples.

However, the following examples are intended to illustrate the present invention, but the present invention is not limited thereto.

Example  1. Recruitment of Alzheimer's patients

First, a total of 115 participants were recruited from the Gil Medical University Gil Medical Center and the Seoul National University Boramae Hospital, 95 and 20, respectively. (2) a mini-mental state examination (MMSE) that is characterized by (1) free recall not associated with or related to other cognitive deficits and not standardized in the initiation signal; (CTL) and 60 biologically confirmed AD patients using clinical dementia rating (CDR). The MMSE is a neurocognitive test designed to screen for memory impairment. This includes scores from 0 to 30, and the higher the score, the better the cognitive abilities. Korean type MMSE is composed of Orientation (10 points), Short term memory and recall (6 points), Attention (5 points), Name waiting (2 points), Command execution (4 points), Judgment (2 points) ). On the other hand, low scores on MMSE indicate that the patient has more severe symptoms of depression. In addition, CDRs are assessed as 0 (no symptoms), 0.5 (mild symptoms), 1 (symptoms of mildness), 2 (moderate symptoms) or 3 (severe symptoms) through structured interviews of subjects and information.

The MMSE score was over 27 for normal subjects aged 65 years and older, and the memory score was normal in MMSE. AD patients aged 65 years or older had an MMSE score of 24 or less and a CDR score of 1 or more. Patients with degenerative neurological diseases such as Parkinson's disease, metabolic disturbances such as cerebrovascular disease, thyroid disease, history of alcohol or drug addiction, trauma history, or neuropsychiatric diseases were excluded from the above patients. The diagnostic criteria for diagnosing AD were DSM-IV of the American Psychiatric Association. All clinical evaluations were performed by the clinical investigator with the subject's genetic status hidden. Characteristics between the evaluated normal and AD patients are shown in Table 1 below. The test was approved by both the Gachon University Road Medical Foundation and the IRB of the Boramae Hospital of Seoul National University.

variable CTL AD Number of participants 55 60 Female (F) / Male (M) 24F / 31M 41F / 19M Average age 70.56 ± 0.65 76.20 ± 0.76 MMSE 27.71 + - 0.17 19.45 ± 0.60 ^** CDR 0.40 + 0.03
(0: n = 10; 0.5: n = 45) 0.78 ± 0.06
(0.5: n = 36; 1: n = 19; 2: n = 5)

^** P < 0.01

As a result, there was no statistically significant difference between MMSE and CDR scores between normal and AD patients as shown in Table 1 (Table 1).

Experimental Example  One. Alzheimer's disease  In patients sNRG -1 expression of the protein

Blood was obtained from 115 participants collected in Test Example 1 in the usual manner. To prevent the coagulation of the obtained blood, heparin was added and centrifuged for 10 minutes at 10,000 占 g at 4 占 폚. After centrifugation, a supernatant corresponding to plasma was obtained, and a protease inhibitor cocktail (Roche) was added thereto and immediately stored at -80 ° C. The stored plasma was prepared by dissolving 1: 200 in PBS buffer and dissolving the day of the experiment.

Immunoblotting was carried out to select proteins whose expression levels were specifically changed in Alzheimer's disease patients in the prepared plasma. Specifically, according to a conventional method, plasma was separated using SDS-PAGE and transferred to a PVDF membrane. The membrane was coated with a primary antibody to the neuregulin-1 (Abcam, UK), APP amyloid beta (Covance, UK), S100a9 (R & D systems, USA), or a control alpha-synuclein (Santa Crus) Were diluted and added at a ratio of 1: 1000, respectively. After addition of HRP (horseradish peroxidase) conjugated secondary antibody (Santa Cruz, USA), protein bands were detected using ECL Plus solution (Amersham Pharmacia, Sweden). The amount of protein was quantified by measuring the intensity of the detected band using Quantity One software 4.6.5 (BioRad, USA). The average expression level is expressed using an arbitrary unit (AU).

As a result, as shown in Fig. 1, all the proteins were detected in plasma samples. However, amyloid beta, S100a9, and alpha synuclein proteins did not show statistically significant differences in expression between normal and AD patients, but the expression level of neurengulin-1 protein was significantly increased in AD patients (FIG. 1).

Experimental Example  2. sNRG -1 and MMSE  Correlation with scores

The correlation between the amount of sNRG-1 protein expressed in Experimental Example 1 and the MMSE score was confirmed by a slot blot method.

In the present study, we used a nonparametric Spearman's rank correlation test (SPSS) in the Statistical Package for the Social Sciences (SPSS) statistical program after matching the MMSE score with the amount of water soluble neuretin-1 (sNRG- Were used to analyze the correlation.

As a result, as shown in FIG. 2, the expression level of sNRG-1 protein showed a negative correlation with the MMSE score (FIG. 2).

Experimental Example  3. Hardness and moderate AD  Between patients sNRG -1 protein expression level

First, AD patients participating in the experiment were classified as mild and moderate patients according to MMSE score or CDR score. At this time, MMSE was classified as middle AD patients with a score of 20 points or less, The expression levels of sNRG-1 protein in the classified patients are shown in Tables 2 and 3, respectively.

variable CTL
(&Gt; 25, n = 55) Longitude AD
(20-24, n = 34) Moderate AD
(6-20, n = 26) P value MMSE 27.71 + 0.17 ^a 22.88 ± 0.23 ^b 14.96 ± 0.68 ^c P < 0.05 sNRG-1 1.00 ± 0.06 ^a 1.26 + - 0.13 ^b 1.32 + 0.14 ^b P < 0.05

variable CTL
(&Lt; 0.5, n = 55) Longitude AD
(0.5, n = 36) Moderate AD
(? 1, n = 24) P value CDR 0.42 ± 0.03 ^a 0.50 ± 0.00 ^b 1.21 0.08 ^c P < 0.05 sNRG-1 1.00 ± 0.06 ^a 1.31 + 0.12 ^b 1.24 ± 0.15 ^b P < 0.05

The other superscripts (a-c) on the same line represent significant differences between groups

As a result, as shown in Table 2, the MMSE score showed a significant difference in the mild and moderate AD patients, and the expression level of sNRG-1 protein was significantly higher in the mild and moderate AD patients than in the normal subjects. However, the expression level of sNRG-1 protein was not significantly different between patients with mild and moderate AD (Table 2).

On the other hand, as shown in Table 3, the sNRG-1 protein expression level did not show any significant difference between the mild and moderate AD patients when classified into the hardness and moderate patients according to the CDR score. However, there were significant differences between patients with mild or moderate AD and normal subjects (Table 3). Therefore, the expression level of sNRG-1 protein was not significantly correlated with the CDR score, but the expression level of sNRG-1 protein was significantly increased in AD patients.

Experimental Example  4. AD  The age of the patient sNRG -1 protein in the rat

First, participants participating in the experiment were classified as 60-69 years old, 70-79 years old, 80 years old or older, and the correlation between sNRG-1 protein expression levels confirmed from the plasma of the patients was shown in FIG. 3

As a result, as shown in Fig. 3, the expression level of sNRG-1 protein of all participants increased little by little as age increased. On the other hand, the expression level of sNRG-1 protein in normal subjects did not show a significant correlation with age, but the expression level of sNRG-1 protein in AD patients was significantly increased with age (Fig. 3).

Experimental Example  5. AD  Patient's sex and sNRG -1 protein in the rat

Participants participating in the experiment were classified according to sex, and the correlation between sNRG-1 protein expression levels confirmed from the plasma of the patients was confirmed, and the results are shown in FIG.

As a result, as shown in FIG. 4, there was no significant difference in the expression amount of sNRG-1 protein according to sex in all participant, normal person, and AD patients (FIG. 4).

<110> Gachon University of Industry-Academic cooperation Foundation          Gil Medical Center          Seoul National University of Industry-Academic cooperation Foundation <120> DIAGNOSTIC COMPOSITION OF ALZHEIMER'S DISEASE USING NEUREGULIN-1 <130> 2016P-12-014 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 220 <212> PRT <213> Artificial Sequence <220> <223> soluble neuregulin-1 <400> 1 Met Ser Glu Arg Lys Glu Gly Arg Gly Lys Gly Lys Gly Lys Lys Lys   1 5 10 15 Asp Arg Gly Ser Arg Gly Lys Pro Ala Pro Gly Glu Gly Asn Pro Ser              20 25 30 Pro Ala Leu Pro Pro Arg Leu Lys Glu Met Lys Ile Gln Glu Ser Ala          35 40 45 Ala Gly Ser Lys Leu Val Leu Arg Cys Glu Thr Ser Ser Glu Tyr Pro      50 55 60 Glu Leu Arg Phe Lys Trp Phe Lys Asn Gly Ser Glu Leu Asn Lys Arg  65 70 75 80 Thr Lys Pro Gln Asn Ile Lys Leu Gln Lys Lys Pro Gly Lys Ser Glu                  85 90 95 Leu Arg Ile Asn Lys Ala Ser Leu Ala Asp Ser Gly Glu Tyr Met Cys             100 105 110 Lys Val Ile Ser Lys Leu Gly Asn Asp Ser Ala Ser Ala Asn Ile Thr         115 120 125 Ile Val Asp Ser Asn Glu Phe Ile Thr Gly Met Pro Ala Ser Thr Glu     130 135 140 Arg Ala Tyr Val Ser Ser Glu Ser Pro Ile Arg Ile Ser Val Ser Thr 145 150 155 160 Glu Gly Ala Asn Thr Ser Ser Thr Ser Thr Ser Thr Thr Gly Thr                 165 170 175 Ser His Leu Val Lys Cys Ala Glu Lys Glu Lys Thr Phe Cys Val Asn             180 185 190 Gly Gly Glu Cys Phe Met Val Lys Asp Leu Ser Asn Pro Ser Arg Tyr         195 200 205 Leu Cys Lys Cys Gln Pro Gly Phe Thr Gly Ala Arg     210 215 220

Claims (10)

A composition for diagnosing Alzheimer's disease comprising an agent for measuring an expression level of a water-soluble neurengulin-1 protein.
delete 2. The composition for diagnosing Alzheimer's disease according to claim 1, wherein the water-soluble neurengulin-1 is a polypeptide represented by SEQ ID NO: 1.
The composition for diagnosing Alzheimer's disease according to claim 1, wherein the preparation is at least one selected from the group consisting of an antibody, an antibody fragment and an aptamer.
The composition for diagnosing Alzheimer's disease according to claim 4, wherein the antibody is any one selected from the group consisting of a monoclonal antibody, a polyclonal antibody and a recombinant antibody.
A kit for diagnosing Alzheimer's disease comprising the composition for diagnosing Alzheimer's disease according to claim 1.
1) measuring the expression level of the water soluble neurengulin-1 protein from the sample; And
2) comparing the amount of water soluble neurengulin-1 protein expressed in step 1) with the amount of soluble neurengulin-1 protein expressed in normal individuals.
delete 8. The method according to claim 7, wherein the sample is at least one selected from the group consisting of urine, blood, serum and plasma.
The method according to claim 7, wherein the expression level is measured by at least one method selected from the group consisting of Western blot, enzyme-immunochemical detection (ELISA), immunohistochemical staining, immunoprecipitation and immunofluorescence. / RTI &gt;

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