JP2017215335A - Biomarker for cognitive impairment disorder and method of detecting cognitive impairment disorder using the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method of detecting cognitive impairment disorder including mild cognitive impairment and Alzheimer's disease using a peptide whose abundances in cognitively unimpaired subjects and cognitively impaired patients are different, and to provide a biomarker, comprising the peptide, for detecting cognitive impairment disorder including mild cognitive impairment and Alzheimer's disease.SOLUTION: Provided are: a biomarker, comprising a transcription factor AP-2 gamma-derived peptide AP2C having an amino acid sequence represented by the sequence number 4, for diagnosing cognitive impairment disorder; and a diagnosis method using the same.SELECTED DRAWING: None

Description

  The present invention relates to biomarkers that are novel proteins and peptides that can be used for detection of cognitive dysfunction diseases including mild cognitive impairment and Alzheimer's disease, and a method for detecting cognitive dysfunction diseases using the biomarkers.

  As a means for discriminating the difference using a sample exhibiting a normal state and a non-normal state of a living body, a technique that has been generally used in an in vitro diagnostic agent is a conventional technique. The most common in vitro diagnostic agents are those that perform diagnostic tests by analyzing blood components as biomarkers. In the prior art in this field, the presence of a single specific protein in blood or a so-called peptide having a molecular weight of 10,000 or less, or the activity in the case of an enzyme protein is measured, and a normal (healthy person) sample and a disease sample are measured. It has helped diagnosis with the obvious difference. That is, measure the amount or activity of single or multiple specific proteins or peptides in a certain number of healthy subjects and samples from disease patients in advance, determine the range of abnormal values and normal values, and evaluate the samples in the same way Measurement is performed and inspection evaluation is performed depending on whether the value belongs to an abnormal value range or a normal value range.

  As a specific measurement method, a specific primary antibody labeled with an enzyme that develops color when a sample is directly or diluted in advance and reacted with a substrate with a single or a plurality of specific proteins or peptides. Enzyme-linked immunosorbent assay (ELISA) or chemiluminescence assay (CLIA) that measures the color development amount of the sample using a secondary antibody and binding to the primary antibody or secondary antibody There are a radioimmunoassay (RIA; RadioImmunoassay) that uses a radioisotope, and an enzyme activity assay that measures the product by color development by directly giving a substrate when the protein is an enzyme. These methods using antibodies will be referred to as enzyme or fluorescent or radioactive substance labeled antibody methods. There is also a method for analyzing enzyme degradation products by high performance liquid chromatography (HPLC). There are also LC-MS / MS method combining HPLC and mass spectrometer and selected reaction monitoring (SRM) / multiple reaction monitoring (MRM) method using this method. After subjecting the sample to appropriate pretreatment, two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) is performed to separate the protein or peptide, and then the target protein or peptide is stained with silver, Coomassie blue, or There is also a method of measuring the concentration in a sample by performing immunostaining (Western blotting) using a corresponding antibody. In addition, there is a technique in which a biological sample is fractionated by column chromatography, and proteins and peptides contained in the fraction are analyzed by mass spectrometry. Further, there is a method of mass spectrometry using a protein chip as a pretreatment instead of column chromatography, and a method of mass spectrometry using magnetic beads as a pretreatment.

  Furthermore, the inventor binds an antibody against a protein or peptide of interest to beads (including magnetic beads), captures the protein or peptide to be measured by this, and then elutes from the beads and measures by mass spectrometry. We are developing. In addition, for the purpose of analyzing an intact protein, a method of performing mass spectrometry using the above method after digestion with trypsin or the like has been reported (see Patent Document 1). However, in any case, utilizing the properties of intact proteins, fractionation is performed as it is, or protein molecules that are specifically adsorbed are selected and analyzed by mass spectrometry.

  Cognitive dysfunction diseases such as Alzheimer's disease are increasing rapidly in Japan with the recent aging. The number was about 1.3 million in 1995, but about 1.9 million in 2005 and is expected to reach about 3 million in 2020. Alzheimer's disease is said to account for 60-90% of cognitive impairment disorders. This disease is not only losing the patient's memory, but also destroying the personality and losing the patient's social life function, so it is becoming a social problem. In Japan, Donepezil hydrochloride, an anti-acetylcholinesterase inhibitor, was approved at the end of 1999, and if it was administered early, it became possible to “delay” the decline in cognitive function with a high probability. In Alzheimer's disease, early diagnosis is the most important issue in order to increase the effects of current therapies and therapeutic drugs to be developed.

The main diagnostic criteria (DSM IV) for Alzheimer's disease by the American Psychiatric Association are shown below.
A. A variety of cognitive deficits manifested by both:
(1) Memory impairment (disability of learning new information or recalling previously learned information)
(2) one or more of the following cognitive impairments
a) Aphasia (language disorder)
b) Apraxia (disability of ability to perform movements despite no impairment of motor function)
c) Agnosia (impaired ability to recognize or identify a subject despite no sensory impairment)
d) Obstacles to ability to execute (plan, organize, order, abstract)
B. Cognitive deficits in criteria A (1) and A (2) each cause a significant impairment in social or occupational function and show a significant decline from the pre-morbid function level (Non-patent Document 1).

  There are various diseases associated with Alzheimer disease (AD). Since dementia such as AD gradually shows a decline in cognitive function, there is a state that should be called a precursor state of dementia. Such a condition is called mild cognitive impairment (MCI). According to US data, 10 to 15% of MCIs who visited a forgetfulness outpatient clinic are transferred to AD in 10 years and approximately 50% in 4 years. Most of the precursor states of AD are included in amnestic MCI. According to the current definition, MCI hears complaints about cognitive decline, but does not interfere with basic daily life. Frontotemporal dementia (FTD) is characterized by cognitive decline and behavior that goes along my way without worrying about the surroundings, as opposed to AD that tries to adjust to the surroundings. FTD includes Pick's disease, which histologically shows the presence of Pick spheres in the cerebral cortex. Lewy body dementia (DLB) is characterized by progressive memory impairment and visual cognitive impairment such as hallucinations. Diagnosis based on clinical symptoms is DLB, accounting for 10-30% of dementia, the second most common degenerative dementia disease in the elderly after (AD). Histologically, it is characterized by the presence of Lewy bodies in the cerebrum. Since FTD and DLB are dementia with dementia, they are also called dementia-type neurological diseases (Non-patent Document 1).

  In the present invention, when collectively referred to as cognitive dysfunction diseases, MCI, AD, and dementia-type neurological diseases are included.

  Tests widely used in the diagnosis of dementia are based on the results of interviews with subjects using the revised Hasegawa Intelligence Scale (HDS-R) and MMSE (Mini-Mental State Examination). . HDS was revised in 1991 and is now called HDS-R. It consists of nine questions and tests for orientation, memorization, computing power, memory / recollection and common sense. Defects are suspected of being 30 out of 30 and below 23. MMSE was devised in the United States for the diagnosis of dementia and covers orientation, memory, computational power, linguistic skills, and graphic skills. It consists of 11 questions with a maximum score of 30. Like HDS-R, it has a score of 23 or less and is suspected of having dementia. The results of both tests agree well with the proportions. These interrogation methods are used only for screening purposes, do not lead to a definitive diagnosis, and neither HDS-R nor MMSE are used for severity classification (Non-patent Document 1).

  The diagnostic imaging includes CT / MRI for cerebral morphological abnormalities such as brain atrophy and cerebral cerebral ventricular enlargement, cerebral blood flow scintigraphy (SPECT) for cerebral blood flow, and oxygen consumption / glucose consumption. There is positron tomography (PET). SPECT and PET are nuclear medicine methods that can detect abnormalities before morphological abnormalities occur (Non-patent Document 1). However, since these image diagnosis requires special equipment, it has a drawback that it cannot be performed in all medical institutions. In addition, judgment may be different depending on the doctor who views the image, and the objectivity is lacking.

  Thus, the diagnosis of dementia, including AD, currently depends on methods that are not objective and require the use of expensive devices, and screening for disease detection is impossible. . If a biomarker is found here that allows objective diagnosis using easily obtained patient samples such as blood (including serum and plasma), screening will present the most important issues It becomes possible to detect early cognitive dysfunction diseases. The present invention provides such a novel biomarker and a method for detecting a cognitive dysfunction disease using the biomarker.

JP 2004-333274 A JP 2006-308533 A

Nakano Imabari, Mizusawa Hidehiro Editing: Alzheimer's Disease, 2004, Nagai Shoten N. Benkirane et al., J. Biol. Chem. Vol. 268, 26279-26285, 1993

  The present invention relates to a non-cognitive impairment subject (including a healthy person, who may be affected by any disease, but who is not affected by a mental disorder including a cognitive impairment disease, age and gender. Detect non-demented control (hereinafter abbreviated as NDC) and cognitive dysfunction patients including cognitive dysfunction diseases including mild cognitive impairment and Alzheimer's disease using proteins and their partial peptides that are present in different amounts The present invention also provides a biomarker for detecting mild cognitive impairment and cognitive dysfunction diseases including Alzheimer's disease comprising the protein and the partial peptide.

  The inventor has intensively studied a method for detecting a cognitive dysfunction disease, and found in the serum a peptide capable of detecting a cognitive dysfunction disease including mild cognitive impairment and Alzheimer's disease. The peptide found in the present invention has significance as a biomarker when it is detected not only in serum but also in other biological samples such as blood, plasma, cerebrospinal fluid, and urine. At the same time, proteins or peptides that are the origin of these peptides (hereinafter referred to as intact proteins or peptides) also have significance as biomarkers.

  Specifically, the present inventor has prepared Complement C3 composed of the amino acid sequence represented by SEQ ID NO: 1, Transcription factor AP-2 gamma composed of the amino acid sequence represented by SEQ ID NO: 3, and the amino acid represented by SEQ ID NO: 5. Synapsin-3 consisting of sequence, Oxytocin receptor consisting of amino acid sequence represented by SEQ ID NO: 7, Inter-alpha-trypsin inhibitor heavy chain H5-like protein consisting of amino acid sequence represented by SEQ ID NO: 9, represented by SEQ ID NO: 11 E3 ubiquitin-protein ligase HERC2 consisting of the amino acid sequence shown below, Prothrombin consisting of the amino acid sequence represented by SEQ ID NO: 13, Transthyretin consisting of the amino acid sequence represented by SEQ ID NO: 15, and the amino acid sequence represented by SEQ ID NO: 17 Tumor necrosis factor receptor superfamily member 16, Complement C4-A consisting of the amino acid sequence represented by SEQ ID NO: 19, amino acid represented by SEQ ID NO: 21 Complement C4-B consisting of a sequence, Fibrinogen alpha chain (isoform 1) consisting of the amino acid sequence represented by SEQ ID NO: 23, and Fibrinogen alpha chain (isoform 2) consisting of the amino acid sequence represented by SEQ ID NO: 25 It has been found that at least one selected protein or peptide or a peptide fragment of 5 or more amino acid residues generated from the protein or peptide can be used as a biomarker for diagnosis of cognitive impairment disorders.

  Further, Complement C3-derived peptide CO3 consisting of the amino acid sequence represented by SEQ ID NO: 2, Transcription factor AP-2 gamma-derived peptide AP2C consisting of the amino acid sequence represented by SEQ ID NO: 4, and the amino acid sequence represented by SEQ ID NO: 6 Synapsin-3 derived peptide SYN3, Oxytocin receptor derived peptide OXYR consisting of the amino acid sequence represented by SEQ ID NO: 8, Inter-alpha-trypsin inhibitor heavy chain H5-like protein derived peptide consisting of the amino acid sequence represented by SEQ ID NO: 10 ITH5L, E3 ubiquitin-protein ligase HERC2-derived peptide HERC2 consisting of the amino acid sequence represented by SEQ ID NO: 12, Prothrombin-derived peptide THRB consisting of the amino acid sequence represented by SEQ ID NO: 14, and amino acid sequence represented by SEQ ID NO: 16 Transthyretin-derived peptide TTHY, Tumor necrosis factor recepto consisting of the amino acid sequence represented by SEQ ID NO: 18 r Superfamily member 16-derived peptide TNR16, Complement C4-derived peptide CO4-1 consisting of the amino acid sequence represented by SEQ ID NO: 20, Complement C4-derived peptide CO4-2 consisting of the amino acid sequence represented by SEQ ID NO: 22, SEQ ID NO: 24 Fibrinogen alpha chain-derived peptide FIBA-1 comprising the amino acid sequence represented, Fibrinogen alpha chain-derived peptide FIBA-2 comprising the amino acid sequence represented by SEQ ID NO: 26, and Fibrinogen alpha comprising the amino acid sequence represented by SEQ ID NO: 27 It has been found that the chain-derived peptide FIBA-3 can be used as a biomarker for diagnosis of cognitive impairment disorders.

  Furthermore, the present inventors have used these proteins and peptides or peptide fragments by using two-dimensional liquid chromatography (2D-LC) MALDI TOF-MS (mass spectrometry) and immunoMS methods, so that many proteins can be obtained at once. In addition, the present inventors have succeeded in measuring peptides or peptide fragments and have completed the present invention.

That is, the aspects of the present invention are as follows.
[1] Complement C3 comprising the amino acid sequence represented by SEQ ID NO: 1, Transcription factor AP-2 gamma comprising the amino acid sequence represented by SEQ ID NO: 3, Synapsin-3 comprising the amino acid sequence represented by SEQ ID NO: 5, Oxytocin receptor consisting of the amino acid sequence represented by SEQ ID NO: 7, Inter-alpha-trypsin inhibitor heavy chain H5-like protein consisting of the amino acid sequence represented by SEQ ID NO: 9, E3 consisting of the amino acid sequence represented by SEQ ID NO: 11 ubiquitin-protein ligase HERC2, Prothrombin consisting of the amino acid sequence represented by SEQ ID NO: 13, Transthyretin consisting of the amino acid sequence represented by SEQ ID NO: 15, Tumor necrosis factor receptor superfamily member 16 consisting of the amino acid sequence represented by SEQ ID NO: 17 Complement C4-A consisting of the amino acid sequence represented by SEQ ID NO: 19, Complement C4-B consisting of the amino acid sequence represented by SEQ ID NO: 21, SEQ ID NO: At least one protein or peptide selected from the group consisting of a fibrinogen alpha chain (isoform 1) consisting of the amino acid sequence represented by 23 and a fibrinogen alpha chain (isoform 2) consisting of the amino acid sequence represented by SEQ ID NO: 25; A biomarker for diagnosis of a cognitive dysfunction disease comprising a peptide fragment having 5 or more amino acid residues generated from the protein or the peptide.

[2] Complement C3-derived peptide CO3 comprising the amino acid sequence represented by SEQ ID NO: 2, Transcription factor AP-2 gamma-derived peptide AP2C comprising the amino acid sequence represented by SEQ ID NO: 4, amino acid sequence represented by SEQ ID NO: 6 Synapsin-3 derived peptide SYN3 consisting of, Oxytocin receptor derived peptide OXYR consisting of the amino acid sequence represented by SEQ ID NO: 8, derived from Inter-alpha-trypsin inhibitor heavy chain H5-like protein consisting of the amino acid sequence represented by SEQ ID NO: 10 Peptide ITH5L, E3 ubiquitin-protein ligase HERC2 derived peptide HERC2 consisting of the amino acid sequence represented by SEQ ID NO: 12, Prothrombin derived peptide THRB consisting of the amino acid sequence represented by SEQ ID NO: 14, from the amino acid sequence represented by SEQ ID NO: 16 Transthyretin derived peptide TTHY, Tumor necrosis factor receptor consisting of amino acid sequence represented by SEQ ID NO: 18 Superfamily member 16-derived peptide TNR16, Complement C4-derived peptide CO4-1 consisting of the amino acid sequence represented by SEQ ID NO: 20, Complement C4-derived peptide CO4-2 consisting of the amino acid sequence represented by SEQ ID NO: 22, and SEQ ID NO: 24 Fibrinogen alpha chain-derived peptide FIBA-1 consisting of the amino acid sequence shown below, Fibrinogen alpha chain-derived peptide FIBA-2 consisting of the amino acid sequence represented by SEQ ID NO: 26, and Fibrinogen alpha chain consisting of the amino acid sequence represented by SEQ ID NO: 27 A biomarker for diagnosis of cognitive dysfunction diseases selected from the derived peptide FIBA-3.

[3] Cognitive dysfunction disease selected from peptides consisting of amino acid sequences represented by SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26 and 27 A biomarker for a cognitive dysfunction disease that appears or increases in a patient biological sample as compared to a biological sample of a subject not suffering from a mental illness.

[4] Alzheimer's disease patient living body selected from peptides consisting of amino acid sequences represented by SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26 and 27 An Alzheimer's disease biomarker that appears or increases in a sample compared to a biological sample of a subject not suffering from a mental illness.

[5] Patients with mild cognitive impairment selected from peptides consisting of amino acid sequences represented by SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26 and 27 A mild cognitive impairment biomarker that appears or increases in a biological sample compared to a biological sample of a subject not suffering from a mental illness.

[6] A method for detecting a cognitive dysfunction disease, comprising measuring at least one biomarker for diagnosing a cognitive dysfunction disease according to any one of [1] to [5] in a biological sample.

[7] Detection is performed by immunoblotting or Western blotting, enzyme or fluorescent or radioactive substance-labeled antibody method, mass spectrometry, immunoMS method, or surface plasmon resonance method. Detection method.

[8] A kit for detecting a cognitive dysfunction disease for measuring at least one biomarker according to any one of [1] to [5].

[9] A detection kit for a cognitive dysfunction disease comprising an antibody or aptamer to at least one biomarker according to any one of [1] to [5].

  According to the present invention, Complement C3 consisting of the amino acid sequence represented by SEQ ID NO: 1 in the biological sample derived from the subject, Transcription factor AP-2 gamma consisting of the amino acid sequence represented by SEQ ID NO: 3, represented by SEQ ID NO: 5 Synapsin-3 consisting of the amino acid sequence, Oxytocin receptor consisting of the amino acid sequence represented by SEQ ID NO: 7, Inter-alpha-trypsin inhibitor heavy chain H5-like protein consisting of the amino acid sequence represented by SEQ ID NO: 9, SEQ ID NO: E3 ubiquitin-protein ligase HERC2 composed of the amino acid sequence represented by 11, Prothrombin composed of the amino acid sequence represented by SEQ ID NO: 13, Transthyretin composed of the amino acid sequence represented by SEQ ID NO: 15, and amino acid represented by SEQ ID NO: 17 Complement C4-A consisting of the amino acid sequence represented by SEQ ID NO: 21, Tumor necrosis factor receptor superfamily member 16 consisting of the sequence, SEQ ID NO: 21 Complement C4-B consisting of the amino acid sequence shown below, Fibrinogen alpha chain (isoform 1) consisting of the amino acid sequence represented by SEQ ID NO: 23, and Fibrinogen alpha chain (isoform 2) consisting of the amino acid sequence represented by SEQ ID NO: 25 Suffering from mental illness by measuring the amount of at least one protein or peptide selected from the group or at least one biomarker selected from peptide fragments of 5 or more amino acid residues arising from the protein or peptide The subject can be diagnosed as suffering from mild cognitive impairment or cognitive dysfunction disease including Alzheimer's disease when it appears or increases compared to the biological sample of the subject who has not.

  Further, according to the present invention, Complement C3-derived peptide CO3 consisting of the amino acid sequence represented by SEQ ID NO: 2 in a biological sample derived from a subject, Transcription factor AP-2 gamma derived from the amino acid sequence represented by SEQ ID NO: 4 Peptide AP2C, Synapsin-3 derived peptide SYN3 comprising the amino acid sequence represented by SEQ ID NO: 6, Oxytocin receptor derived peptide OXYR comprising the amino acid sequence represented by SEQ ID NO: 8, Inter comprising the amino acid sequence represented by SEQ ID NO: 10. -alpha-trypsin inhibitor heavy chain H5-like protein-derived peptide ITH5L, E3 ubiquitin-protein ligase HERC2-derived peptide HERC2 consisting of the amino acid sequence represented by SEQ ID NO: 12, Prothrombin-derived peptide consisting of the amino acid sequence represented by SEQ ID NO: 14 THRB, transthyretin-derived peptide TTHY consisting of the amino acid sequence represented by SEQ ID NO: 16, amino acid represented by SEQ ID NO: 18 Tumor necrosis factor receptor superfamily member 16-derived peptide TNR16 consisting of an acid sequence, Complement C4-derived peptide CO4-1 consisting of the amino acid sequence represented by SEQ ID NO: 20, Complement C4-derived peptide CO4 consisting of the amino acid sequence represented by SEQ ID NO: 22 -2, Fibrinogen alpha chain-derived peptide FIBA-1 consisting of the amino acid sequence represented by SEQ ID NO: 24, Fibrinogen alpha chain-derived peptide FIBA-2 consisting of the amino acid sequence represented by SEQ ID NO: 26, and SEQ ID NO: 27 By measuring the type and amount of at least one peptide selected from the group consisting of the fibrinogen alpha chain-derived peptide FIBA-3 consisting of the amino acid sequence described above, it is compared with that in a biological sample of a subject not suffering from mental illness. If it appears or increases, the subject may have mild cognitive impairment or cognitive impairment, including Alzheimer's disease. It can be diagnosed as affected.

  The present invention also provides a diagnostic system that is both extremely accurate and specific. According to the present invention, it becomes possible for the first time to make a highly accurate diagnosis for a cognitive dysfunction disease for which there is no specific test method for biological samples such as blood. Furthermore, the biomarker of the present invention is highly useful in determining drug effects.

The point in the rectangle shown by (A) (the group is called a cluster) is the retention time (retention time) of the mass peak in C18 reverse phase chromatography (2nd dimension) of Marker A detected in the serum of each subject. And m / z. The points in the cluster can be regarded as having the same m / z and the same retention time within the error range, and are defined as originating from the same peptide. Difference analysis results for Marker A (dot diagram). Marker A is a complement C3-derived peptide CO3 as shown in the amino acid sequence obtained as a result of the MS / MS analysis in FIG. FIG. 2 shows a comparison between CO3 and cognitive impairment disorders (AD, MCI, DLB, FTD) and NDC. SEQ ID NO: 2 ROC curve of CO3. See the Examples and Results section for ROC curve definitions. A) is an AD vs. NDC, and B) is an ROC curve obtained by analysis with a receiver operating characteristic curve in a comparison of MCI vs. NDC. MS / MS spectrum diagram by TOF / TOF mass spectrometer. The upper part of FIG. 4 shows the amino acid sequence of CO3 and b ions and y ions appearing in the MS / MS spectrum. The result of a difference analysis of SEQ ID NO: 4 AP2C. Comparison between subjects without mental illness (NDC) and cognitive impairment patients (AD, MCI, DLB, FTD). The result of a difference analysis of SEQ ID NO: 6 SYN3. Comparison between subjects without mental illness (NDC) and cognitive impairment patients (AD, MCI, DLB, FTD). The result of a difference analysis of sequence number 8 OXYR. Comparison between subjects without mental illness (NDC) and cognitive impairment patients (AD, MCI, DLB, FTD). The result of a difference analysis of SEQ ID NO: 10 ITH5L. Comparison between subjects without mental illness (NDC) and cognitive impairment patients (AD, MCI, DLB, FTD). The result of a difference analysis of SEQ ID NO: 12 HERC2. Comparison between subjects without mental illness (NDC) and cognitive impairment patients (AD, MCI, DLB, FTD). SEQ ID NO: 14 Result of THRB difference analysis. Comparison between subjects without mental illness (NDC) and cognitive impairment patients (AD, MCI, DLB, FTD). The result of a difference analysis of SEQ ID NO: 16 TTHY. Comparison between subjects without mental illness (NDC) and cognitive impairment patients (AD, MCI, DLB, FTD). The result of a difference analysis of SEQ ID NO: 18 TNR16. Comparison between subjects without mental illness (NDC) and cognitive impairment patients (AD, MCI, DLB, FTD). The result of a difference analysis of SEQ ID NO: 20 CO4-1. Comparison between subjects without mental illness (NDC) and cognitive impairment patients (AD, MCI, DLB, FTD). The result of a difference analysis of SEQ ID NO: 22 CO4-2. Comparison between subjects without mental illness (NDC) and cognitive impairment patients (AD, MCI, DLB, FTD). The result of a difference analysis of SEQ ID NO: 24 FIBA-1. Comparison between subjects without mental illness (NDC) and cognitive impairment patients (AD, MCI, DLB, FTD). The result of a difference analysis of SEQ ID NO: 26 FIBA-2. Comparison between subjects without mental illness (NDC) and cognitive impairment patients (AD, MCI, DLB, FTD). The result of a difference analysis of SEQ ID NO: 27 FIBA-3. Comparison between subjects without mental illness (NDC) and cognitive impairment patients (AD, MCI, DLB, FTD).

  The present invention detects the type and amount of an intact protein and / or its partial peptide and simultaneously measures the variation of the type and amount of the intact protein and its partial peptide when the subject suffers from a cognitive impairment disease. This is a method for diagnosing whether or not a subject suffers from a cognitive dysfunction disease. Here, the peptide generally refers to a peptide in which amino acids having a molecular weight of 10,000 or less are linked, or the number of amino acid residues is from several to about 50 or less. In the present invention, a partial peptide of an intact protein can be used as a biomarker for detecting a cognitive dysfunction disease. In the case of a partial peptide, a peptide having a partial amino acid sequence of the amino acid sequence of the intact protein. In this case, the molecular weight is 10,000 or less. In the present invention, the intact peptide partial peptide refers to a peptide having a partial amino acid sequence of the amino acid sequence of the intact protein, and is generated as a partial peptide in the expression synthesis process by transcription and translation; After being synthesized as an intact protein, it may be digested and decomposed in vivo to produce a digested degradation product peptide. This is because protein synthesis and control mechanisms are deregulated when the living body is in a state other than normal, such as a cognitive impairment disorder. That is, the present invention determines whether a subject is in a normal state or suffers from a cognitive dysfunction disease by using expression synthesis and / or digestion degradation of protein in vivo as an index, and also suffers from a cognitive dysfunction disease. It is also a method for evaluating and discriminating the degree of progression of the disease in the case of In the present invention, the detection of a cognitive dysfunction disease refers to evaluation discrimination, that is, diagnosis, whether or not a subject suffers from cognitive dysfunction disease. It may also include an assessment of the risk that the subject will suffer from more severe cognitive impairment.

  In the method of the present invention, as an intact protein that can be used as a biomarker for detecting a cognitive impairment disorder, specifically, Complement C3 consisting of the amino acid sequence represented by SEQ ID NO: 1, represented by SEQ ID NO: 3 Transcription factor AP-2 gamma composed of amino acid sequence, Synapsin-3 composed of amino acid sequence represented by SEQ ID NO: 5, Oxytocin receptor composed of amino acid sequence represented by SEQ ID NO: 7, and amino acid sequence represented by SEQ ID NO: 9 Inter-alpha-trypsin inhibitor heavy chain H5-like protein, E3 ubiquitin-protein ligase HERC2 consisting of the amino acid sequence represented by SEQ ID NO: 11, Prothrombin consisting of the amino acid sequence represented by SEQ ID NO: 13, represented by SEQ ID NO: 15 Transthyretin consisting of the amino acid sequence shown, and Tumor necrosis factor receptor superfamily consisting of the amino acid sequence shown in SEQ ID NO: 17. Complement C4-A comprising the amino acid sequence represented by member 16, SEQ ID NO: 19, Complement C4-B comprising the amino acid sequence represented by SEQ ID NO: 21, Fibrinogen alpha chain comprising the amino acid sequence represented by SEQ ID NO: 23 ( isoform 1) and fibrinogen alpha chain (isoform 2) consisting of the amino acid sequence represented by SEQ ID NO: 25, and peptide fragments of 5 or more amino acid residues which are partial peptides of these intact proteins have the same purpose. Can be used.

  Further, as partial peptides that can be used as biomarkers for detecting cognitive impairment disorders, Complement C3-derived peptide CO3 consisting of the amino acid sequence represented by SEQ ID NO: 2 and Transcription factor AP consisting of the amino acid sequence represented by SEQ ID NO: 4 -2 gamma derived peptide AP2C, Synapsin-3 derived peptide SYN3 consisting of amino acid sequence represented by SEQ ID NO: 6, Oxytocin receptor derived peptide OXYR consisting of amino acid sequence represented by SEQ ID NO: 8, amino acid represented by SEQ ID NO: 10 Inter-alpha-trypsin inhibitor heavy chain peptide HTH-like protein derived from sequence ITH5L, E3 ubiquitin-protein ligase HERC2-derived peptide HERC2 composed of amino acid sequence represented by SEQ ID NO: 12, and amino acid sequence represented by SEQ ID NO: 14 Prothrombin-derived peptide THRB, transthyretin-derived peptide consisting of the amino acid sequence represented by SEQ ID NO: 16 Peptide TTHY, Tumor necrosis factor receptor superfamily member 16-derived peptide TNR16 consisting of the amino acid sequence represented by SEQ ID NO: 18, Complement C4-derived peptide CO4-1 consisting of the amino acid sequence represented by SEQ ID NO: 20, and SEQ ID NO: 22 Complement C4-derived peptide CO4-2 consisting of an amino acid sequence, Fibrinogen alpha chain-derived peptide FIBA-1 consisting of the amino acid sequence represented by SEQ ID NO: 24, Fibrinogen alpha chain-derived peptide FIBA consisting of the amino acid sequence represented by SEQ ID NO: 26 And Fibrinogen alpha chain-derived peptide FIBA-3 consisting of the amino acid sequence represented by -2 and SEQ ID NO: 27. In the present invention, the above intact protein and peptide are used as a marker. The amino acid sequence represented by SEQ ID NOs: 1-27 consists of an amino acid sequence in which one or several amino acids are deleted, substituted, or added. Proteins and peptides are also included, and these proteins or peptides can also be used as biomarkers in the methods of the invention. Here, “one or several” means “one or three”, “one or two” or “one”. Furthermore, these peptides that can be used as biomarkers for detecting cognitive dysfunction diseases also include peptide fragments having 5 or more amino acid residues resulting from the amino acid sequence represented by SEQ ID NOs: 1 to 27. The reason why the peptide fragment is 5 or more amino acid residues is due to the following description in Non-Patent Document 2. That is, the peptide CGGGERA in which R is replaced with K for the amino acid residue sequence IRGERA at the C-terminus (130-135) of histone H3 and IR is deleted, and CGG is bound to GERA instead. It was reported that it was recognized by the antibody obtained. This indicates that antigenic recognition is made by a peptide consisting of 4 or more amino acid residues. In the present invention, in order to have generality other than the C-terminal of histone H3, the number of residues is increased by 1 to 5 or more, but such low molecular peptides are also targeted. It is important when using a method for detection and fractionation using immunological techniques such as immunoblotting, ELISA, and immunoMS.

  A sugar chain may be added to an intact protein or a partial peptide thereof. Proteins and partial peptides to which these sugar chains are added can also be used as biomarkers for detecting cognitive impairment disorders.

  In the present invention, the biomarker may be quantified or the presence or absence may be determined by qualitative.

  As a method for separating a biomarker in a biological sample such as serum in the present invention, two-dimensional electrophoresis or two-dimensional chromatography can be used. The two types of chromatography in this case may be selected from known chromatography such as ion exchange chromatography, reverse phase chromatography, gel filtration chromatography and the like. It can also be quantified by the SRM / MRM method using the LC-MS / MS method. Furthermore, an immunoMS that binds an antibody against the protein or peptide of interest to the beads (including magnetic beads) developed by the inventor, captures the protein or peptide to be measured, and then elutes from the bead and measures by mass spectrometry By using this method, the presence or amount of the target protein, protein fragment, or peptide can be easily evaluated without using two-dimensional electrophoresis or chromatography.

  According to the method of the present invention, a subject's cognitive dysfunction can be evaluated at a mild stage, which is useful for preventive medicine. Furthermore, when psychotherapy or pharmacotherapy is performed on a patient suffering from a cognitive dysfunction disease, if the progression of the disorder is suppressed, it is also reflected in the amount of protein / partial peptide in a biological sample such as serum. By measuring this, it is possible to evaluate and determine the therapeutic effect.

  The type and amount of protein in the biological sample can be measured by various methods. When a target protein (including a protein fragment and a partial peptide) has been identified and an antibody against the protein (primary antibody) has been obtained, the following method can be used.

1. Immunoblot method The simplest method. Prepare test serum diluted in several stages, drop a certain amount (around 1 microliter) onto a suitable membrane such as nitrocellulose membrane, and air dry. After treatment with a blocking solution containing a protein such as BSA, washing and reacting with the primary antibody, and after washing, a labeled secondary antibody for detecting the primary antibody is reacted. After washing the membrane, the label is visualized and the concentration is measured.

2. Western blotting After one- or two-dimensional gel electrophoresis including isoelectric point or SDS-PAGE, the separated protein is temporarily transferred to a suitable membrane such as PVDF membrane and labeled with the primary antibody. Using the prepared secondary antibody, the abundance of the target protein is measured in the same manner as in the immunoblotting method described above.

3. ELISA method An antibody against a protein or its partial peptide is bound to a carrier such as a microtiter plate that has been subjected to special chemical modification in advance. After serial dilution of the sample, an appropriate amount is added to the microtiter plate to which the antibody is bound and incubated. . Thereafter, washing is performed to remove uncaptured proteins and partial peptides. Next, a secondary antibody conjugated with a fluorescent or chemiluminescent substance or enzyme is added and incubated. For detection, evaluation is performed by adding each substrate and then measuring visible light by fluorescent or chemiluminescent substances or enzymatic reactions. A substance capable of binding to a protein or a partial peptide thereof may be used instead of an antibody. For example, an aptamer or the like can be used.

Furthermore, although a method (refer patent document 2) is illustrated below, it is not limited to them.
4). Method using microarray (microchip) A microarray is a generic term for devices in which substances that can be bound to a substance to be measured are aligned (array) immobilized on a carrier (substrate). In the case of the present invention, antibodies or aptamers against proteins and partial peptides may be used after being aligned and immobilized. For measurement, add a biological sample to the immobilized antibody, etc., bind the protein or partial peptide to be measured on the microarray, and then add the secondary antibody to which the fluorescent or chemiluminescent substance or enzyme is bound. Incubate. Detection may be performed by adding each substrate and then measuring the fluorescent or chemiluminescent substance or visible light from the enzyme reaction.

5. Mass Spectrometry In mass spectrometry, for example, an antibody against a specific protein and its partial peptide is bound to microbeads or substrates (protein chips) that have been specially modified in advance. The microbead may be a magnetic bead. Any material can be used for the substrate. The antibodies to be used are (1) an antibody that recognizes only the full length of a specific protein, (2) an antibody that recognizes only a partial peptide, (3) an antibody that recognizes both a specific protein and its partial peptide, or A combination of (1) and (2), (1) and (3), or (2) and (3) may be used. A sample is serially diluted with a stock solution or a buffer solution, and an appropriate amount thereof is added to a microbead or a substrate to which an antibody is bound, and incubated. Thereafter, washing is performed to remove uncaptured proteins and partial peptides. Then, the proteins and partial peptides captured on the microbeads or the substrate are analyzed by mass spectrometry using MALDI-TOF-MS, SELDI-TOF-MS, etc. Measure peak intensity. A certain amount of an appropriate internal standard substance is added to the original biological sample, its peak intensity is measured, and the ratio to the peak intensity of the target substance is determined to obtain the concentration in the original biological sample. I can know. This method is called immunoMS method. Further, the sample can be diluted with a stock solution or a buffer solution or a part of protein can be removed, and then separated by HPLC and quantified by mass spectrometry using an electrospray ionization (ESI) method. At that time, the concentration in the sample can be known by absolute quantification by the SRM / MRM method using an isotope-labeled internal standard peptide.

  Furthermore, in addition to the above methods, proteins and partial peptides can be analyzed by a method using two-dimensional electrophoresis, a method using surface plasmon resonance, or the like.

  The present invention also includes a method in which a biological sample collected from a subject is subjected to two-dimensional electrophoresis or surface plasmon resonance, and a cognitive impairment disease is detected using the presence or amount of the biomarker as an index.

  Search for marker peptides for diagnosis of cognitive dysfunction by two-dimensional liquid chromatography-mass spectrometry (2D-LC MALDI TOF-MS)

(1) Serum sample Hereinafter, the parentheses are abbreviations.
40 AD (Alzheimer's disease), 35 MCI (mild cognitive impairment), 13 DLB (Lewy body dementia), 7 FTD (frontotemporal dementia), and NDC (not suffering from mental illness) Subject) 21 subjects.

(2) Method 475 μl of 0.1% trifluoroacetic acid (TFA) was added to each 25 μl of serum and boiled at 100 ° C. for 15 minutes. Thereafter, in order to recover a peptide having a molecular weight of 10,000 or less, ultrafiltration was performed using Millipore YM-10. Next, analysis by the 2D-LC MALDI TOF-MS method was performed as follows. That is, samples collected by ultrafiltration were fractionated into a total of 382 per sample using two-dimensional HPLC (first dimension SCX cation exchange and second dimension C18 reverse phase column). In the SCX cation exchange column, the fraction was divided into two fractions, that is, SCX 1 was a flow-through fraction and SCX 2 was a fraction eluted at a salt concentration of 100%. The two fractions fractionated by SCX were fractionated into 191 fractions by C18 reverse phase column chromatography, respectively. One fraction eluted in 6 seconds, and the retention time (Retention Time) was determined by multiplying the number of eluted fractions minus 1 by 6 seconds. All samples are MALDI target plates (MTP AnchorChip (TM) 600/384) for MALDI TOF / TOF mass spectrometer (ultraflex TOF / TOF, BRUKER DALTONICS) using an online spotting robot (AccuSpot, SHIMADZU) The wells on the plate (BRUKER DALTONICS) were spotted and co-crystallized while mixing with the matrix solution (α-cyano-hydroxycinnamic acid, α-CHCA). After mounting on ultraflex TOF / TOF, laser was irradiated and the mass and peak area at that mass (including the normalization process described below) were automatically measured in reflectron mode. The peak area was normalized with 250 fmole of bradykinin 1-7 per well previously added to the matrix solution to obtain an Area value. That is, the area value was obtained by dividing the peak area at a specific mass of the sample by the peak area obtained from bradykinin 1-7. This Area value corresponds to 25 μl of sample serum. Detection of peptides (difference analysis) with differences in serum abundance between groups was performed using Parnassum (TM) (MCBI), a multigroup statistical analysis software developed by us. Peptides in which a difference in abundance was observed were determined by MS / MS using ultraflex TOF / TOF, and the intact protein or peptide that originated the peptide was identified.

(3) Results The following shows the results of differential analysis using Parnassum software for the data obtained by subjecting each subject's serum to 2D-LC MALDI TOF-MS. Figure 1 shows that the first fraction (SCX 1) of the two fractions obtained from the first-dimension SCX cation exchange column is 191 fractions on the second-dimension C18 reversed-phase column. In the spectrum measured by MALDI TOF-MS, the horizontal axis represents the mass per unit charge (m / z), and the vertical axis represents the retention time of the second-phase reversed-phase column chromatography. FIG. Each point in the figure is a TOF-MS peak derived from an individual subject, and the part where the points are collected is a peak derived from one peptide having the same m / z and the same retention time within the error range. It is defined and shown as a cluster. The set of points in the rectangle shown in FIG. 1A is a Marker A cluster.

  FIG. 2 shows the result of the difference analysis of Marker A. Marker A is a complement C3-derived peptide CO3 as shown later in FIG. FIG. 2 is a comparison between cognitive impairment disorders (AD, MCI, DLB, FTD) and subjects not suffering from mental disorders (NDC). Area values were statistically significantly higher (p <0.05) in cognitive impairment disorders (AD, MCI, DLB, FTD) as a result of t-test compared to NDC.

From the results of FIG. 2, in order to evaluate how useful Marker A is as a biomarker, an analysis using a receiver operating characteristic curve (ROC curve) was performed. A) and B) in FIG. 3 are ROC curves in comparison of AD vs. NDC and MCI vs. NDC, respectively. The closer the area value (AUC of ROC) (hereinafter referred to as AUC value) of the ROC curve is to 1, the higher the usefulness as a biomarker. Typical values of sensitivity and specificity are A) and B) in Fig. 3.When a straight line is drawn from the 100% point on the y-axis to the ROC curve, the ROC that minimizes the distance It is the value of the point coordinates (open square in the figure) on the curve. The cut-off value giving this point is a useful threshold for classification between different groups, and the sensitivity and specificity (that is, the above-mentioned representative values) at that time are indicators of the usefulness of the biomarker together with the AUC value.
In FIG. 3A), sensitivity as a typical value in AD vs NDC is 73.0%, specificity is 100%, AUC value is 0.88, and B) is typical in MCI vs NDC. The sensitivity (sensitivity) was 70.6%, the specificity was 89.5%, and the AUC value was 0.83.

  These results indicate that Marker A is useful for distinguishing AD and MCI patients from non-psychiatric subjects (NDC). In particular, since MCI is a pre-stage of AD, Marker A is considered to be extremely useful as a marker for early diagnosis and detection of subjects who may transition to AD.

  FIG. 4 shows the result of MS / MS analysis of Marker A using an ultraflex TOF / TOF type mass spectrometer. Signals indicating y-ion, b-ion, and a-ion appeared sufficiently, and the amino acid sequence could be easily determined. A search by Mascot was performed on this result, and the protein or peptide that was the origin (hereinafter referred to as the intact protein or peptide) was identified as Complement C3, and it was found that the detected peptide was the amino acid sequence APVIHQEMIGGLRN derived from this protein. It became. For detected peptides, UniProt Entry Name CO3 is used as an abbreviation for peptide names. The same applies to peptides other than CO3.

  Peptides with different abundance in serum between groups, including Marker A, were subjected to MS / MS with ultraflex TOF / TOF to determine the amino acid sequence and identify the intact protein or peptide. The results are shown below. For peptides other than Marker A, signals indicating y-ion and b-ion appeared sufficiently, and the amino acid sequence could be easily determined. The following amino acid sequences show two sets of sequences as one set, the first of which is the intact protein amino acid sequence and the second is the sequence of the detected peptide. The underlined portion of the first sequence corresponds to the detected peptide sequence. The amino acid sequence starting from (0001) in the sequence represents the N-terminal side.

[1] Complement C3-derived peptide CO3
CO3 of SEQ ID NO: 2 formed the cluster of FIG. 1 by clustering using Parnassum (TM). As shown in FIG. 2, the Area value is statistically significantly higher (p <0.05) in the cognitive dysfunction disease (AD, MCI, DLB, FTD) than the NDC, as a result of the t-test, It has been found that CO3 of SEQ ID NO: 2 is useful in distinguishing between patients with cognitive dysfunction disease (AD, MCI, DLB, FTD) and subjects not suffering from mental illness (NDC). Analysis by the receiver operating characteristic curve clearly shows that it is useful for distinguishing AD and MCI patients from subjects not suffering from mental illness (FIG. 3A), B), Table 1. ).
Intact protein / peptide
0001 SPMYSIITPN ILRLESEETM VLEAHDAQGD VPVTVTVHDF PGKKLVLSSE
0051 KTVLTPATNH MGNVTFTIPA NREFKSEKGR NKFVTVQATF GTQVVEKVVL
0101 VSLQSGYLFI QTDKTIYTPG STVLYRIFTV NHKLLPVGRT VMVNIENPEG
0151 IPVKQDSLSS QNQLGVLPLS WDIPELVNMG QWKIRAYYEN SPQQVFSTEF
0201 EVKEYVLPSF EVIVEPTEKF YYIYNEKGLE VTITARFLYG KKVEGTAFVI
0251 FGIQDGEQRI SLPESLKRIP IEDGSGEVVL SRKVLLDGVQ NPRAEDLVGK
0301 SLYVSATVIL HSGSDMVQAE RSGIPIVTSP YQIHFTKTPK YFKPGMPFDL
0351 MVFVTNPDGS PAYRVPVAVQ GEDTVQSLTQ GDGVAKLSIN THPSQKPLSI
0401 TVRTKKQELS EAEQATRTMQ ALPYSTVGNS NNYLHLSVLR TELRPGETLN
0451 VNFLLRMDRA HEAKIRYYTY LIMNKGRLLK AGRQVREPGQ DLVVLPLSIT
0501 TDFIPSFRLV AYYTLIGASG QREVVADSVW VDVKDSCVGS LVVKSGQSED
0551 RQPVPGQQMT LKIEGDHGAR VVLVAVDKGV FVLNKKNKLT QSKIWDVVEK
0601 ADIGCTPGSG KDYAGVFSDA GLTFTSSSGQ QTAQRAELQC PQPAARRRRS
0651 VQLTEKRMDK VGKYPKELRK CCEDGMRENP MRFSCQRRTR FISLGEACKK
0701 VFLDCCNYIT ELRRQHARAS HLGLARSNLD EDIIAEENIV SRSEFPESWL
0751 WNVEDLKEPP KNGISTKLMN IFLKDSITTW EILAVSMSDK KGICVADPFE
0801 VTVMQDFFID LRLPYSVVRN EQVEIRAVLY NYRQNQELKV RVELLHNPAF
0851 CSLATTKRRH QQTVTIPPKS SLSVPYVIVP LKTGLQEVEV KAAVYHHFIS
0901 DGVRKSLKVV PEGIRMNKTV AVRTLDPERL GREGVQKEDI PPADLSDQVP
0951 DTESETRILL QGTPVAQMTE DAVDAERLKH LIVTPSGCGE QNMIGMTPTV
1001 IAVHYLDETE QWEKFGLEKR QGALELIKKG YTQQLAFRQP SSAFAAFVKR
1051 APSTWLTAYV VKVFSLAVNL IAIDSQVLCG AVKWLILEKQ KPDGVFQED A
1101 PVIHQEMIGG LRN NNEKDMA LTAFVLISLQ EAKDICEEQV NSLPGSITKA
1151 GDFLEANYMN LQRSYTVAIA GYALAQMGRL KGPLLNKFLT TAKDKNRWED
1201 PGKQLYNVEA TSYALLALLQ LKDFDFVPPV VRWLNEQRYY GGGYGSTQAT
1251 FMVFQALAQY QKDAPDHQEL NLDVSLQLPS RSSKITHRIH WESASLLRSE
1301 ETKENEGFTV TAEGKGQGTL SVVTMYHAKA KDQLTCNKFD LKVTIKPAPE
1351 TEKRPQDAKN TMILEICTRY RGDQDATMSI LDISMMTGFA PDTDDLKQLA
1401 NGVDRYISKY ELDKAFSDRN TLIIYLDKVS HSEDDCLAFK VHQYFNVELI
1451 QPGAVKVYAY YNLEESCTRF YHPEKEDGKL NKLCRDELCR CAEENCFIQK
1501 SDDKVTLEER LDKACEPGVD YVYKTRLVKV QLSNDFDEYI MAIEQTIKSG
1551 SDEVQVGQQR TFISPIKCRE ALKLEEKKHY LMWGLSSDFW GEKPNLSYII
1601 GKDTWVEHWP EEDECQDEEN QKQCQDLGAF TESMVVFGCP N (SEQ ID NO: 1)

Complement C3-derived peptide CO3
APVIHQEMIG GLRN (SEQ ID NO: 2)

[2] Transcription factor AP-2 gamma-derived peptide AP2C
The area value of AP2C of SEQ ID NO: 4 is statistically significantly (p <0.05) higher in cognitive dysfunction diseases (AD, MCI, DLB, FTD) as a result of t-test compared to NDC (Fig. 5) It has been found that AP2C of SEQ ID NO: 4 is useful in distinguishing between patients with cognitive impairment disorders (AD, MCI, DLB, FTD) and subjects not suffering from mental disorders (NDC). Analysis by recipient operating characteristic curves clearly proves useful in distinguishing AD and MCI patients from subjects not suffering from mental illness (Table 1).
Intact protein / peptide
0001 MLWKITDNVK YEEDCEDRHD GSSNGNPRVP HLSSAGQHLY SPAPPLSHTG
0051 VAEYQPPPYF PPPYQQLAYS QSADPYSHLG EAYAAAINPL HQPAPTGSQQ
0101 QAW PGRQSQE GAGLPSHHG R PAGLLPHLSG LEAGAVSARR DAYRRSDLLL
0151 PHAHALDAAG LAENLGLHDM PHQMDEVQNV DDQHLLLHDQ TVIRKGPISM
0201 TKNPLNLPCQ KELVGAVMNP TEVFCSVPGR LSLLSSTSKY KVTVAEVQRR
0251 LSPPECLNAS LLGGVLRRAK SKNGGRSLRE KLDKIGLNLP AGRRKAAHVT
0301 LLTSLVEGEA VHLARDFAYV CEAEFPSKPV AEYLTRPHLG GRNEMAARKN
0351 MLLAAQQLCK EFTELLSQDR TPHGTSRLAP VLETNIQNCL SHFSLITHGF
0401 GSQAICAAVS ALQNYIKEAL IVIDKSYMNP GDQSPADSNK TLEKMEKHRK (SEQ ID NO: 3)

Transcription factor AP-2 gamma-derived peptide AP2C
PGRQSQEGAG LPSHHG (SEQ ID NO: 4)

[3] Synapsin-3 derived peptide SYN3
The area value of SYN3 of SEQ ID NO: 6 is statistically significantly (p <0.05) higher in cognitive dysfunction diseases (AD, MCI, DLB, FTD) compared to NDC as a result of t-test (Fig. 6) It was found that SYN3 of SEQ ID NO: 6 is useful in distinguishing between patients with cognitive impairment disorders (AD, MCI, DLB, FTD) and subjects who do not suffer from mental disorders (NDC). Analysis by recipient operating characteristic curves clearly proves useful in distinguishing AD and MCI patients from subjects not suffering from mental illness (Table 1).
Intact protein / peptide
0001 MNFLRRRLSD SSFMANLPNG YMTDLQRPDS STSSPASPAM ERRHPQPLAA
0051 SFSSPGSSLF SSLSSAMKQA PQATSGLMEP PGPSTPIVQR PRILLVIDDA
0101 HTDWSKYFHG KKVNGEIEIR VEQAEFSELN LAAYVTGGCM VDMQVVRNGT
0151 KVVSRSFKPD FILVRQHAYS MALGEDYRSL VIGLQYGGLP AVNSLYSVYN
0201 FCSKPWVFSQ LIKIFHSLGP EKFPLVEQTF FPNHKPMVTA PHFPVVVKLG
0251 HAHAGMGKIK VENQLDFQDI TSVVAMAKTY ATTEAFIDSK YDIRIQKIGS
0301 NYKAYMRTSI SGNWKANTGS AMLEQVAMTE RYRLWVDSCS EMFGGLDICA
0351 VKAVHSK DGR DYIIEVMDSS MPLIGEHVEE DRQLMADLVV SKMSQLPMPG
0401 GTAPSPLRPW APQIKSAKSP GQAQLGPQLG QPQPRPPPQG GPRQAQSPQP
0451 QRSGSPSQQR LSPQGQQPLS PQSGSPQQQR SPGSPQLSRA SSGSSPNQAS
0501 KPGATLASQP RPPVQGRSTS QQGEESKKPA PPHPHLNKSQ SLTNSLSTSD
0551 TSQRGTPSED EAKAETIRNL RKSFASLFSD (SEQ ID NO: 5)

Synapsin-3 derived peptide SYN3
EMFGGLDICA VKAVHSK (SEQ ID NO: 6)

[4] Oxytocin receptor-derived peptide OXYR
The area value of OXYR of SEQ ID NO: 8 is statistically significantly (p <0.05) higher in cognitive dysfunction diseases (AD, MCI, DLB, FTD) than in NDC as a result of t-test (Fig. 7) The OXYR of SEQ ID NO: 8 was found to be useful in distinguishing between patients with cognitive dysfunction disorders (AD, MCI, DLB, FTD) and subjects not suffering from mental disorders (NDC). Analysis by recipient operating characteristic curves clearly proves useful in distinguishing AD and MCI patients from subjects not suffering from mental illness (Table 1).
Intact protein / peptide
0001 MEGALAANWS AEAANAS AAP PGAEGNRT AG PPRRNEALAR VEVAVLCLIL
0051 LLALSGNACV LLALRTTRQK HSRLFFFMKH LSIADLVVAV FQVLPQLLWD
0101 ITFRFYGPDL LCRLVKYLQV VGMFASTYLL LLMSLDRCLA ICQPLRSLRR
0151 RTDRLAVLAT WLGCLVASAP QVHIFSLREV ADGVFDCWAV FIQPWGPKAY
0201 ITWITLAVYI VPVIVLAACY GLISFKIWQN LRLKTAAAAA AEAPEGAAAG
0251 DGGRVALARV SSVKLISKAK IRTVKMTFII VLAFIVCWTP FFFVQMWSVW
0301 DANAPKEASA FIIVMLLASL NSCCNPWIYM LFTGHLFHEL VQRFLCCSAS
0351 YLKGRRLGET SASKKSNSSS FVLSHRSSSQ RSCSQPSTA (SEQ ID NO: 7)

Oxytocin receptor-derived peptide OXYR
AAPPGAEGNR T (SEQ ID NO: 8)

[5] Inter-alpha-trypsin inhibitor heavy chain peptide derived from H5-like protein ITH5L
The area value of ITH5L of SEQ ID NO: 10 is statistically significantly (p <0.05) higher in cognitive dysfunction diseases (AD, MCI, DLB) than in NDC as a result of t-test (FIG. 8). It has been found that ITH5L of SEQ ID NO: 10 is useful in distinguishing between patients with cognitive dysfunction disease (AD, MCI, DLB) and subjects not suffering from mental illness (NDC). Analysis by recipient operating characteristic curves clearly proves useful in distinguishing AD and MCI patients from subjects not suffering from mental illness (Table 1).
Intact protein / peptide
0001 GPPVPASSST KLLMTSYSMR STVVSRYAHT LVTSVLFNPH AEAHEAIFDL
0051 DLPHLAFISN FTMTINNKVY IAEVKEKHQA KKIYEEAHQQ GKTAAHVGIR
0101 DRESEKFRIS TSLAAGTEVT FSLAYEELLQ RHQGQYQLVV SLRPGQLVKR
0151 LSIEVTVSER TGISYVHIPP LRTGRLRTNA HASEVDSPPS TRIERGETCV
0201 RITYCPTLQD QSSISGSGIM ADFLVQYDVV MEDIIGDVQI YDDYFIHYFA
0251 PRGLPPMEKN VVFVIDVSSS MFGTKMEQTK TAMNVILSDL QANDYFNIIS
0301 FSDTVNVWKA GGSIQATIQN VHSAKDYLHC MEADGWTDVN SALLAAASVL
0351 NHSNQEPGRG PSVGRIPLII FLTDGEPTAG VTTPSVILSN VRQALGH RVS
0401 LFSLAFGDDA D FTLLRRLSL ENRGIARRIY EDTDAALQLK GLYEEISMPL
0451 LADVRLNYLG GLVGASPWAV FPNYFGGSEL VVAGQVQPGK QELGIHLAAR
0501 GPKDQLLVAH HSEGATNNSQ KAFGCPGEPA PNVAHFIRRL WAYVTIGELL
0551 DAHFQARDTT TRHLLAAKVL NLSLEYNFVT PLTSLVMVQP KQASEETRRQ
0601 TSTSAGPDTI MPSSSSRHGL GVSTAQPALV PKVISPKSRP VKPKFYLSST
0651 TTASTKKMLS SKELEPLGES PHTLSMPTYP KAKIPAQQDS GTLAQPTLRT
0701 KPTILVPSNS GTLLPLKPGS LSHQNPDILP TNSRTQVPPV KPGIPASPKA
0751 DTVKCVTPLH SKPGAPSHPQ LGALTSQAPK GLPQSRPGVS TLQVPKYPLH
0801 TRPRVPAPKT RNNMPHLGPG ILLSKTPKIL LSLKPSAPPH QISTSISLSK
0851 PETPNPHMPQ TPLPPRPDRP RPPLPESLST FPNTISSSTG PSSTTTTSVL
0901 GEPLPMPFTP TLPPGRFWHQ YDLLPGPQRT RQVLGPSRPG VPTMSLLNSS
0951 RPTPEGSPPN LPILLPSSIL PEAISLLLLP EELELLSESM VESKFVESLN
1001 PPAFYTFLTP DEDGSPNWDG NSEEILGGAG GSMESQGSSV GLAKGTLPSI
1051 FTFSSSVDGD PHFVIQIPHS EEKICFTLNG HPGDLLQLIE DPKAGLHVSG
1101 KLLGAPPRPG HKDQTRTYFQ IITVTTDKPR AYTITISRSS ISLRGEGTLR
1151 LSWDQPALLK RPQLELYVAA AARLTLRLGP YLEFLVLRHR YRHPSTLQLP
1201 HLGFYVANGS GLSPSARGLI GQFQHADIRL VTGPMGPCLR RHHGPDVPVI
1251 LGKRLLKDSP RLLPRWASCW LVKRSHVELL LGHPYLSYVL (SEQ ID NO: 9)

Inter-alpha-trypsin inhibitor heavy chain H5-like protein derived peptide ITH5L
RVSLFSLAFG DDAD (SEQ ID NO: 10)

[6] E3 ubiquitin-protein ligase HERC2-derived peptide HERC2
The area value of HERC2 of SEQ ID NO: 12 is statistically significantly (p <0.05) higher in cognitive dysfunction diseases (AD, MCI, DLB) as a result of t-test than in NDC (FIG. 9). The HERC2 of SEQ ID NO: 12 was found to be useful in distinguishing between patients with cognitive dysfunction disease (AD, MCI, DLB) and subjects who do not suffer from mental illness (NDC). Analysis by recipient operating characteristic curves clearly proves useful in distinguishing AD and MCI patients from subjects not suffering from mental illness (Table 1).
Intact protein / peptide
0001 MPSESFCLAA QARLDSKWLK TDIQLAFTRD GLCGLWNEMV KDGEIVYTGT
0051 ESTQNGELPP RKDDSVEPSG TKKEDLNDKE KKDEEETPAP IYRAKSILDS
0101 WVWGKQPDVN ELKECLSVLV KEQQALAVQS ATTTLSALRL KQRLVILERY
0151 FIALNRTVFQ ENVKVKWKSS GISLPPVDKK SSRPAGKGVE GLARVGSRAA
0201 LSFAFAFLRR AWRSGEDADL CSELLQESLD ALRALPEASL FDESTVSSVW
0251 LEVVERATRF LRSVVTGDVH GTPATKGPGS IPLQDQHLAL AILLELAVQR
0301 GTLSQMLSAI LLLLQLWDSG AQETDNERSA QGTSAPLLPL LQRFQSIICR
0351 KDAPHSEGDM HLLSGPLSPN ESFLRYLTLP QDNELAIDLR QTAVVVMAHL
0401 DRLATPCMPP LCSSPTSHKG SLQEVIGWGL IGWKYYANVI GPIQCEGLAN
0451 LGVTQIACAE KRFLILSRNG RVYTQAYNSD TLAPQLVQGL ASRNIVKIAA
0501 HSDGHHYLAL AATGEVYSWG CGDGGRLGHG DTVPLEEPKV ISAFSGKQAG
0551 KHVVHIACGS TYSAAITAEG ELYTWGRGNY GRLGHGSSED EAIPMLVAGL
0601 KGLKVIDVAC GSGDAQTLAV TENGQVWSWG DGDYGKLGRG GSDGCKTPKL
0651 IEKLQDLDVV KVRCGSQFSI ALTKDGQVYS WGKGDNQRLG HGTEEHVRYP
0701 KLLEGLQGKK VIDVAAGSTH CLALTEDSEV HSWGSNDQCQ HFDTLRVTKP
0751 EPAALPGLDT KHIVGIACGP AQSFAWSSCS EWSIGLRVPF VVDICSMTFE
0801 QLDLLLRQVS EGMDGSADWP PPQEKECVAV ATLNLLRLQL HAAISHQVDP
0851 EFLGLGLGSI LLNSLKQTVV TLASSAGVLS TVQSAAQAVL QSGWSVLLPT
0901 AEERARALSA LLPCAVSGNE VNISPGRRFM IDLLVGSLMA DGGLESALHA
0951 AITAEIQDIE AKKEAQKEKE IDEQEANAST FHRSRTPLDK DLINTGICES
1001 SGKQCLPLVQ LIQQLLRNIA SQTVARLKDV ARRISSCLDF EQHSRERSAS
1051 LDLLLRFQRL LISKLYPGES IGQTSDISSP ELMGVGSLLK KYTALLCTHI
1101 GDILPVAASI ASTSWRHFAE VAYIVEGDFT GVLLPELVVS IVLLLSKNAG
1151 LMQEAGAVPL LGGLLEHLDR FNHLAPGKER DDHEELAWPG IMESFFTGQN
1201 CRNNEEVTLI RKADLENHNK DGGFWTVIDG KVYDIKDFQT QSLTGNSILA
1251 QFAGEDPVVA LEAALQFEDT RESMHAFCVG QYLEPDQEIV TIPDLGSLSS
1301 PLIDTERNLG LLLGLHASYL AMSTPLSPVE IECAKWLQSS IFSGGLQTSQ
1351 IHYSYNEEKD EDHCSSPGGT PASKSRLCSH RRALGDHSQA FLQAIADNNI
1401 QDHNVKDFLC QIERYCRQCH LTTPIMFPPE HPVEEVGRLL LCCLLKHEDL
1451 GHVALSLVHA GALGIEQVKH RTLPKSVVDV CRVVYQAKCS LIKTHQEQGR
1501 SYKEVCAPVI ERLRFLFNEL RPAVCNDLSI MSKFKLLSSL PRWRRIAQKI
1551 IRERRKKRVP KKPESTDDEE KIGNEESDLE EACILPHSPI NVDKRPIAIK
1601 SPKDKWQPLL STVTGVHKYK WLKQNVQGLY PQSPLLSTIA EFALKEEPVD
1651 VEKMRKCLLK QLERAEVRLE GIDTILKLAS KNFLLPSVQY AMFCGWQRLI
1701 PEGIDIGEPL TDCLKDVDLI PPFNRMLLEV TFGKLYAWAV QNIRNVLMDA
1751 SAKFKELGIQ PVPLQTITNE NPSGPSLGTI PQARFLLVML SMLTLQHGAN
1801 NLDLLLNSGM LALTQTALRL IGPSCDNVEE DMNASAQGAS ATVLEETRKE
1851 TAPVQLPVSG PELAAMMKIG TRVMRGVDWK WGDQDGPPPG LGRVIGELGE
1901 DGWIRVQWDT GSTNSYRMGK EGKYDL KLAE LPAAAQPSAE DSD TEDDSEA
1951 EQTERNIHPT AMMFTSTINL LQTLCLSAGV HAEIMQSEAT KTLCGLLRML
2001 VESGTTDKTS SPNRLVYREQ HRSWCTLGFV RSIALTPQVC GALSSPQWIT
2051 LLMKVVEGHA PFTATSLQRQ ILAVHLLQAV LPSWDKTERA RDMKCLVEKL
2101 FDFLGSLLTT CSSDVPLLRE STLRRRRVRP QASLTATHSS TLAEEVVALL
2151 RTLHSLTQWN GLINKYINSQ LRSITHSFVG RPSEGAQLED YFPDSENPEV
2201 GGLMAVLAVI GGIDGRLRLG GQVMHDEFGE GTVTRITPKG KITVQFSDMR
2251 TCRVCPLNQL KPLPAVAFNV NNLPFTEPML SVWAQLVNLA GSKLEKHKIK
2301 KSTKQAFAGQ VDLDLLRCQQ LKLYILKAGR ALLSHQDKLR QILSQPAVQE
2351 TGTVHTDDGA VVSPDLGDMS PEGPQPPMIL LQQLLASATQ PSPVKAIFDK
2401 QELEAAALAV CQCLAVESTH PSSPGFEDCS SSEATTPVAV QHIRPARVKR
2451 RKQSPVPALP IVVQLMEMGF SRRNIEFALK SLTGASGNAS SLPGVEALVG
2501 WLLDHSDIQV TELSDADTVS DEYSDEEVVE DVDDAAYSMS TGAVVTESQT
2551 YKKRADFLSN DDYAVYVREN IQVGMMVRCC RAYEEVCEGD VGKVIKLDRD
2601 GLHDLNVQCD WQQKGGTYWV RYIHVELIGY PPPSSSSHIK IGDKVRVKAS
2651 VTTPKYKWGS VTHQSVGVVK AFSANGKDII VDFPQQSHWT GLLSEMELVP
2701 SIHPGVTCDG CQMFPINGSR FKCRNCDDFD FCETCFKTKK HNTRHTFGRI
2751 NEPGQSAVFC GRSGKQLKRC HSSQPGMLLD SWSRMVKSLN VSSSVNQASR
2801 LIDGSEPCWQ SSGSQGKHWI RLEIFPDVLV HRLKMIVDPA DSSYMPSLVV
2851 VSGGNSLNNL IELKTININP SDTTVPLLND CTEYHRYIEI AIKQCRSSGI
2901 DCKIHGLILL GRIRAEEEDL AAVPFLASDN EEEEDEKGNS GSLIRKKAAG
2951 LESAATIRTK VFVWGLNDKD QLGGLKGSKI KVPSFSETLS ALNVVQVAGG
3001 SKSLFAVTVE GKVYACGEAT NGRLGLGISS GTVPIPRQIT ALSSYVVKKV
3051 AVHSGGRHAT ALTVDGKVFS WGEGDDGKLG HFSRMNCDKP RLIEALKTKR
3101 IRDIACGSSH SAALTSSGEL YTWGLGEYGR LGHGDNTTQL KPKMVKVLLG
3151 HRVIQVACGS RDAQTLALTD EGLVFSWGDG DFGKLGRGGS EGCNIPQNIE
3201 RLNGQGVCQI ECGAQFSLAL TKSGVVWTWG KGDYFRLGHG SDVHVRKPQV
3251 VEGLRGKKIV HVAVGALHCL AVTDSGQVYA WGDNDHGQQG NGTTTVNRKP
3301 TLVQGLEGQK ITRVACGSSH SVAWTTVDVA TPSVHEPVLF QTARDPLGAS
3351 YLGVPSDADS SAASNKISGA SNSKPNRPSL AKILLSLDGN LAKQQALSHI
3401 LTALQIMYAR DAVVGALMPA AMIAPVECPS FSSAAPSDAS AMASPMNGEE
3451 CMLAVDIEDR LSPNPWQEKR EIVSSEDAVT PSAVTPSAPS ASARPFIPVT
3501 DDLGAASIIA ETMTKTKEDV ESQNKAAGPE PQALDEFTSL LIADDTRVVV
3551 DLLKLSVCSR AGDRGRDVLS AVLSGMGTAY PQVADMLLEL CVTELEDVAT
3601 DSQSGRLSSQ PVVVESSHPY TDDTSTSGTV KIPGAEGLRV EFDRQCSTER
3651 RHDPLTVMDG VNRIVSVRSG REWSDWSSEL RIPGDELKWK FISDGSVNGW
3701 GWRFTVYPIM PAAGPKELLS DRCVLSCPSM DLVTCLLDFR LNLASNRSIV
3751 PRLAASLAAC AQLSALAASH RMWALQRLRK LLTTEFGQSI NINRLLGEND
3801 GETRALSFTG SALAALVKGL PEALQRQFEY EDPIVRGGKQ LLHSPFFKVL
3851 VALACDLELD TLPCCAETHK WAWFRRYCMA SRVAVALDKR TPLPRLFLDE
3901 VAKKIRELMA DSENMDVLHE SHDIFKREQD EQLVQWMNRR PDDWTLSAGG
3951 SGTIYGWGHN HRGQLGGIEG AKVKVPTPCE ALATLRPVQL IGGEQTLFAV
4001 TADGKLYATG YGAGGRLGIG GTESVSTPTL LESIQHVFIK KVAVNSGGKH
4051 CLALSSEGEV YSWGEAEDGK LGHGNRSPCD RPRVIESLRG IEVVDVAAGG
4101 AHSACVTAAG DLYTWGKGRY GRLGHSDSED QLKPKLVEAL QGHRVVDIAC
4151 GSGDAQTLCL TDDDTVWSWG DGDYGKLGRG GSDGCKVPMK IDSLTGLGVV
4201 KVECGSQFSV ALTKSGAVYT WGKGDYHRLG HGSDDHVRRP RQVQGLQGKK
4251 VIAIATGSLH CVCCTEDGEV YTWGDNDEGQ LGDGTTNAIQ RPRLVAALQG
4301 KKVNRVACGS AHTLAWSTSK PASAGKLPAQ VPMEYNHLQE IPIIALRNRL
4351 LLLHHLSELF CPCIPMFDLE GSLDETGLGP SVGFDTLRGI LISQGKEAAF
4401 RKVVQATMVR DRQHGPVVEL NRIQVKRSRS KGGLAGPDGT KSVFGQMCAK
4451 MSSFGPDSLL LPHRVWKVKF VGESVDDCGG GYSESIAEIC EELQNGLTPL
4501 LIVTPNGRDE SGANRDCYLL SPAARAPVHS SMFRFLGVLL GIAIRTGSPL
4551 SLNLAEPVWK QLAGMSLTIA DLSEVDKDFI PGLMYIRDNE ATSEEFEAMS
4601 LPFTVPSASG QDIQLSSKHT HITLDNRAEY VRLAINYRLH EFDEQVAAVR
4651 EGMARVVPVP LLSLFTGYEL ETMVCGSPDI PLHLLKSVAT YKGIEPSASL
4701 IQWFWEVMES FSNTERSLFL RFVWGRTRLP RTIADFRGRD FVIQVLDKYN
4751 PPDHFLPESY TCFFLLKLPR YSCKQVLEEK LKYAIHFCKS IDTDDYARIA
4801 LTGEPAADDS SDDSDNEDVD SFASDSTQDY LTGH (SEQ ID NO: 11)

E3 ubiquitin-protein ligase HERC2-derived peptide HERC2
KLAELPAAAQ PSAEDSD (SEQ ID NO: 12)

[7] Prothrombin-derived peptide THRB
The area value of THRB of SEQ ID NO: 14 is statistically significantly (p <0.05) higher in cognitive dysfunction diseases (AD, MCI, DLB, FTD) as a result of t-test compared to NDC (Fig. 10) The THRB of SEQ ID NO: 14 was found to be useful in distinguishing between patients with cognitive impairment disorders (AD, MCI, DLB, FTD) and subjects not suffering from mental disorders (NDC). Analysis by recipient operating characteristic curves clearly proves useful in distinguishing AD and MCI patients from subjects not suffering from mental illness (Table 1).
Intact protein / peptide
0001 ANTFLEEVRK GNLERECVEE TCSYEEAFEA LESSTATDVF WAKYTACETA
0051 RTPRDKLAAC LEGNCAEGLG TNYRGHVNIT RSGIECQLWR SRYPHKPEIN
0101 STTHPGADLQ ENFCRNPDSS TTGPWCYTTD PTVRRQECSI PVCGQDQVTV
0151 AMTPRSEGSS VNLSPPLEQC VPDRGQQYQG RLAVTTHGLP CLAWASAQAK
0201 ALSKHQDFNS AVQLVENFCR NPDGDEEGVW CYVAGKPGDF GYCDLNYCEE
0251 AVEEETGDGL DEDSDRAIEG R TATSEYQTF FNPRTFGSGE AD CGLRPLFE
0301 KKSLEDKTER ELLESYIDGR IVEGSDAEIG MSPWQVMLFR KSPQELLCGA
0351 SLISDRWVLT AAHCLLYPPW DKNFTENDLL VRIGKHSRTR YERNIEKISM
0401 LEKIYIHPRY NWRENLDRDI ALMKLKKPVA FSDYIHPVCL PDRETAASLL
0451 QAGYKGRVTG WGNLKETWTA NVGKGQPSVL QVVNLPIVER PVCKDSTRIR
0501 ITDNMFCAGY KPDEGKRGDA CEGDSGGPFV MKSPFNNRWY QMGIVSWGEG
0551 CDRDGKYGFY THVFRLKKWI QKVIDQFGE (SEQ ID NO: 13)

Prothrombin-derived peptide THRB
TATSEYQTFF NPRTFGSGEA D (SEQ ID NO: 14)

[8] Transthyretin-derived peptide TTHY
The area value of TTHY of SEQ ID NO: 16 is statistically significantly (p <0.05) higher in cognitive dysfunction diseases (AD, MCI, DLB, FTD) as a result of t-test compared to NDC (Fig. 11) It was found that TTHY of SEQ ID NO: 16 is useful in distinguishing between patients with cognitive impairment disorders (AD, MCI, DLB, FTD) and subjects not suffering from mental disorders (NDC). Analysis by recipient operating characteristic curves clearly proves useful in distinguishing AD and MCI patients from subjects not suffering from mental illness (Table 1).
Intact protein / peptide
0001 GPTGTGESKC PLMVKVLD AV RGSPAINVAV HVFRKAAD DT WEPFASGKTS
0051 ESGELHGLTT EEEFVEGIYK VEIDTKSYWK ALGISPFHEH AEVVFTANDS
0101 GPRRYTIAAL LSPYSYSTTA VVTNPKE (SEQ ID NO: 15)

Transthyretin-derived peptide TTHY
AVRGSPAINV AVHVFRKAAD (SEQ ID NO: 16)

[9] Tumor necrosis factor receptor superfamily member 16-derived peptide TNR16
The area value of TNR16 of SEQ ID NO: 18 is statistically significantly (p <0.05) higher in cognitive impairment disorders (AD, MCI, DLB, FTD) as a result of t-test compared to NDC (Fig. 12) It was found that TNR16 of SEQ ID NO: 18 is useful in distinguishing between patients with cognitive impairment disorders (AD, MCI, DLB, FTD) and subjects not suffering from mental disorders (NDC). Analysis by recipient operating characteristic curves clearly proves useful in distinguishing AD and MCI patients from subjects not suffering from mental illness (Table 1).
Intact protein / peptide
0001 KEACPTGLYT HSGECCKACN LGEGVAQPCG ANQTVCEPCL DSVTFSDVVS
0051 ATEPCKPCTE CVGLQSMSAP CVEADDAVCR CAYGYYQDET TGRCEACRVC
0101 EAGSGLVFSC QDKQNTVCEE CPDGTYSDEA NHVDPCLPCT VCEDTERQLR
0151 ECTRWADAEC EEIPGRWITR STPPEGSDST APSTQEPEAP PEQDLIASTV
0201 AGVVTTVMGS SQPVVTRGTT DNLIPVYCSI LAAVVVGLVA YIAFKRWNSC
0251 KQNKQGANSR PVNQTPPPEG EKLHSDSGIS VDSQSLHDQQ PHT QTASGQA
0301 LKGDGGLYS S LPPAKREEVE KLLNGSAGDT WRHLAGELGY QPEHIDSFTH
0351 EACPVRALLA SWATQDSATL DALLAALRRI QRADLVESLC SESTATSPV (SEQ ID NO: 17)

Tumor necrosis factor receptor superfamily member 16-derived peptide TNR16
QTASGQALKG DGGLYS (SEQ ID NO: 18)

[10] Complement C4-A derived peptide CO4-1
The area value of CO4-1 of SEQ ID NO: 20 is statistically significantly (p <0.05) higher in cognitive impairment disorders (AD, MCI, DLB, FTD) as a result of t-test than NDC (FIG. 13), CO4-1 of SEQ ID NO: 20 is useful in distinguishing between patients with cognitive dysfunction disorders (AD, MCI, DLB, FTD) and subjects not suffering from mental disorders (NDC) I understood. Analysis by recipient operating characteristic curves clearly proves useful in distinguishing AD and MCI patients from subjects not suffering from mental illness (Table 1).
Complement C4-A protein is separated into Complement C4 beta chain, Complement C4-A alpha chain and Complement C4 gamma chain by biosynthesis and processing. SEQ ID NO: 19 is the amino acid sequence of intact protein Complement C4-A, which includes all of these.
Intact protein / peptide
0001 KPRLLLFSPS VVHLGVPLSV GVQLQDVPRG QVVKGSVFLR NPSRNNVPCS
0051 PKVDFTLSSE RDFALLSLQV PLKDAKSCGL HQLLRGPEVQ LVAHSPWLKD
0101 SLSRTTNIQG INLLFSSRRG HLFLQTDQPI YNPGQRVRYR VFALDQKMRP
0151 STDTITVMVE NSHGLRVRKK EVYMPSSIFQ DDFVIPDISE PGTWKISARF
0201 SDGLESNSST QFEVKKYVLP NFEVKITPGK PYILTVPGHL DEMQLDIQAR
0251 YIYGKPVQGV AYVRFGLLDE DGKKTFFRGL ESQTKLVNGQ SHISLSKAEF
0301 QDALEKLNMG ITDLQGLRLY VAAAIIESPG GEMEEAELTS WYFVSSPFSL
0351 DLSKTKRHLV PGAPFLLQAL VREMSGSPAS GIPVKVSATV SSPGSVPEVQ
0401 DIQQNTDGSG QVSIPIIIPQ TISELQLSVS AGSPHPAIAR LTVAAPPSGG
0451 PGFLSIERPD SRPPRVGDTL NLNLRAVGSG ATFSHYYYMI LSRGQIVFMN
0501 REPKRTLTSV SVFVDHHLAP SFYFVAFYYH GDHPVANSLR VDVQAGACEG
0551 KLELSVDGAK QYRNGESVKL HLETDSLALV ALGALDTALY AAGSKSHKPL
0601 NMGKVFEAMN SYDLGCGPGG GDSALQVFQA AGLAFSDGDQ WTLSRKRLSC
0651 PKEKTTRKKR NVNFQKAINE KLGQYASPTA KRCCQDGVTR LPMMRSCEQR
0701 AARVQQPDCR EPFLSCCQFA ESLRKKSRDK GQAGLQRALE ILQEEDLIDE
0751 DDIPVRSFFP ENWLWRVETV DRFQILTLWL PDSLTTWEIH GLSLSKTKGL
0801 CVATPVQLRV FREFHLHLRL PMSVRRFEQL ELRPVLYNYL DKNLTVSVHV
0851 SPVEGLCLAG GGGLAQQVLV PAGSARPVAF SVVPTAAAAV SLKVVARGSF
0901 EFPVGDAVSK VLQIEKEGAI HREELVYELN PLDHRGRTLE IPGNSDPNMI
0951 PDGDFNSYVR VTASDPLDTL GSEGALSPGG VASLLRLPRG CGEQTMIYLA
1001 PTLAASRYLD KTEQWSTLPP ETKDHAVDLI QKGYMRIQQF RKADGSYAAW
1051 LSRDSSTWLT AFVLKVLSLA QEQVGGSPEK LQETSNWLLS QQQADGSFQD
1101 PCPVLDRSMQ GGLVGNDETV ALTAFVTIAL HHGLAVFQDE GAEPLKQRVE
1151 ASISKANSFL GEKASAGLLG AHAAAITAYA LSLTKAPVDL LGVAHNNLMA
1201 MAQETGDNLY WGSVTGSQSN AVSPTPAPRN PSDPMPQAPA LWIETTAYAL
1251 LHLLLHEGKA EMADQASAWL TRQGSFQGGF RSTQDTVIAL DALSAYWIAS
1301 HTTEERGLNV TLSSTGR NGF KSHALQLNNR QIR GLEEELQ FSLGSKINVK
1351 VGGNSKGTLK VLRTYNVLDM KNTTCQDLQI EVTVKGHVEY TMEANEDYED
1401 YEYDELPAKD DPDAPLQPVT PLQLFEGRRN RRRREAPKVV EEQESRVHYT
1451 VCIWRNGKVG LSGMAIADVT LLSGFHALRA DLEKLTSLSD RYVSHFETEG
1501 PHVLLYFDSV PTSRECVGFE AVQEVPVGLV QPASATLYDY YNPERRCSVF
1551 YGAPSKSRLL ATLCSAEVCQ CAEGKCPRQR RALERGLQDE DGYRMKFACY
1601 YPRVEYGFQV KVLREDSRAA FRLFETKITQ VLHFTKDVKA AANQMRNFLV
1651 RASCRLRLEP GKEYLIMGLD GATYDLEGHP QYLLDSNSWI EEMPSERLCR
1701 STRQRAACAQ LNDFLQEYGT QGCQV (SEQ ID NO: 19)

Complement C4-derived peptide CO4-1
NGFKSHALQL NNRQIR (SEQ ID NO: 20)

[11] Complement C4-B derived peptide CO4-1
Peptide CO4-1 of SEQ ID NO: 20 is an amino acid sequence present in the topological region portion common to the proteins of Complement C4-A (SEQ ID NO: 19) and Complement C4-B based on the results of MS / MS and MASCOT database search. .
Complement C4-B protein is divided into Complement C4 beta chain, Complement C4-B alpha chain, and Complement C4 gamma chain by processing after biosynthesis. SEQ ID NO: 21 is the amino acid sequence of intact protein Complement C4-B, which includes all of these.
Intact protein / peptide
0001 KPRLLLFSPS VVHLGVPLSV GVQLQDVPRG QVVKGSVFLR NPSRNNVPCS
0051 PKVDFTLSSE RDFALLSLQV PLKDAKSCGL HQLLRGPEVQ LVAHSPWLKD
0101 SLSRTTNIQG INLLFSSRRG HLFLQTDQPI YNPGQRVRYR VFALDQKMRP
0151 STDTITVMVE NSHGLRVRKK EVYMPSSIFQ DDFVIPDISE PGTWKISARF
0201 SDGLESNSST QFEVKKYVLP NFEVKITPGK PYILTVPGHL DEMQLDIQAR
0251 YIYGKPVQGV AYVRFGLLDE DGKKTFFRGL ESQTKLVNGQ SHISLSKAEF
0301 QDALEKLNMG ITDLQGLRLY VAAAIIESPG GEMEEAELTS WYFVSSPFSL
0351 DLSKTKRHLV PGAPFLLQAL VREMSGSPAS GIPVKVSATV SSPGSVPEVQ
0401 DIQQNTDGSG QVSIPIIIPQ TISELQLSVS AGSPHPAIAR LTVAAPPSGG
0451 PGFLSIERPD SRPPRVGDTL NLNLRAVGSG ATFSHYYYMI LSRGQIVFMN
0501 REPKRTLTSV SVFVDHHLAP SFYFVAFYYH GDHPVANSLR VDVQAGACEG
0551 KLELSVDGAK QYRNGESVKL HLETDSLALV ALGALDTALY AAGSKSHKPL
0601 NMGKVFEAMN SYDLGCGPGG GDSALQVFQA AGLAFSDGDQ WTLSRKRLSC
0651 PKEKTTRKKR NVNFQKAINE KLGQYASPTA KRCCQDGVTR LPMMRSCEQR
0701 AARVQQPDCR EPFLSCCQFA ESLRKKSRDK GQAGLQRALE ILQEEDLIDE
0751 DDIPVRSFFP ENWLWRVETV DRFQILTLWL PDSLTTWEIH GLSLSKTKGL
0801 CVATPVQLRV FREFHLHLRL PMSVRRFEQL ELRPVLYNYL DKNLTVSVHV
0851 SPVEGLCLAG GGGLAQQVLV PAGSARPVAF SVVPTAAAAV SLKVVARGSF
0901 EFPVGDAVSK VLQIEKEGAI HREELVYELN PLDHRGRTLE IPGNSDPNMI
0951 PDGDFNSYVR VTASDPLDTL GSEGALSPGG VASLLRLPRG CGEQTMIYLA
1001 PTLAASRYLD KTEQWSTLPP ETKDHAVDLI QKGYMRIQQF RKADGSYAAW
1051 LSRDSSTWLT AFVLKVLSLA QEQVGGSPEK LQETSNWLLS QQQADGSFQD
1101 LSPVIHRSMQ GGLVGNDETV ALTAFVTIAL HHGLAVFQDE GAEPLKQRVE
1151 ASISKANSFL GEKASAGLLG AHAAAITAYA LSLTKAPVDL LGVAHNNLMA
1201 MAQETGDNLY WGSVTGSQSN AVSPTPAPRN PSDPMPQAPA LWIETTAYAL
1251 LHLLLHEGKA EMADQASAWL TRQGSFQGGF RSTQDTVIAL DALSAYWIAS
1301 HTTEERGLNV TLSSTGR NGF KSHALQLNNR QIR GLEEELQ FSLGSKINVK
1351 VGGNSKGTLK VLRTYNVLDM KNTTCQDLQI EVTVKGHVEY TMEANEDYED
1401 YEYDELPAKD DPDAPLQPVT PLQLFEGRRN RRRREAPKVV EEQESRVHYT
1451 VCIWRNGKVG LSGMAIADVT LLSGFHALRA DLEKLTSLSD RYVSHFETEG
1501 PHVLLYFDSV PTSRECVGFE AVQEVPVGLV QPASATLYDY YNPERRCSVF
1551 YGAPSKSRLL ATLCSAEVCQ CAEGKCPRQR RALERGLQDE DGYRMKFACY
1601 YPRVEYGFQV KVLREDSRAA FRLFETKITQ VLHFTKDVKA AANQMRNFLV
1651 RASCRLRLEP GKEYLIMGLD GATYDLEGHP QYLLDSNSWI EEMPSERLCR
1701 STRQRAACAQ LNDFLQEYGT QGCQV (SEQ ID NO: 21)

As a precaution, the sequence of CO4-1 is shown below.
Complement C4-derived peptide CO4-1
NGFKSHALQL NNRQIR (SEQ ID NO: 20)

[12] Complement C4-A derived peptide CO4-2
The area value of CO4-2 of SEQ ID NO: 22 is statistically significantly (p <0.05) higher in cognitive dysfunction diseases (AD, MCI, DLB, FTD) than in NDC as a result of t-test. (FIG. 14), CO4-2 of SEQ ID NO: 22 is useful in distinguishing between patients with cognitive dysfunction disorders (AD, MCI, DLB, FTD) and subjects who do not suffer from mental disorders (NDC) I understood. Analysis by recipient operating characteristic curves clearly proves useful in distinguishing AD and MCI patients from subjects not suffering from mental illness (Table 1).
Complement C4-A protein is separated into Complement C4 beta chain, Complement C4-A alpha chain and Complement C4 gamma chain by biosynthesis and processing. SEQ ID NO: 19 is the amino acid sequence of intact protein Complement C4-A, which includes all of these.
Intact protein / peptide
0001 KPRLLLFSPS VVHLGVPLSV GVQLQDVPRG QVVKGSVFLR NPSRNNVPCS
0051 PKVDFTLSSE RDFALLSLQV PLKDAKSCGL HQLLRGPEVQ LVAHSPWLKD
0101 SLSRTTNIQG INLLFSSRRG HLFLQTDQPI YNPGQRVRYR VFALDQKMRP
0151 STDTITVMVE NSHGLRVRKK EVYMPSSIFQ DDFVIPDISE PGTWKISARF
0201 SDGLESNSST QFEVKKYVLP NFEVKITPGK PYILTVPGHL DEMQLDIQAR
0251 YIYGKPVQGV AYVRFGLLDE DGKKTFFRGL ESQTKLVNGQ SHISLSKAEF
0301 QDALEKLNMG ITDLQGLRLY VAAAIIESPG GEMEEAELTS WYFVSSPFSL
0351 DLSKTKRHLV PGAPFLLQAL VREMSGSPAS GIPVKVSATV SSPGSVPEVQ
0401 DIQQNTDGSG QVSIPIIIPQ TISELQLSVS AGSPHPAIAR LTVAAPPSGG
0451 PGFLSIERPD SRPPRVGDTL NLNLRAVGSG ATFSHYYYMI LSRGQIVFMN
0501 REPKRTLTSV SVFVDHHLAP SFYFVAFYYH GDHPVANSLR VDVQAGACEG
0551 KLELSVDGAK QYRNGESVKL HLETDSLALV ALGALDTALY AAGSKSHKPL
0601 NMGKVFEAMN SYDLGCGPGG GDSALQVFQA AGLAFSDGDQ WTLSRKRLSC
0651 PKEKTTRKKR NVNFQKAINE KLGQYASPTA KRCCQDGVTR LPMMRSCEQR
0701 AARVQQPDCR EPFLSCCQFA ESLRKKSRDK GQAGLQRALE ILQEEDLIDE
0751 DDIPVRSFFP ENWLWRVETV DRFQILTLWL PDSLTTWEIH GLSLSKTKGL
0801 CVATPVQLRV FREFHLHLRL PMSVRRFEQL ELRPVLYNYL DKNLTVSVHV
0851 SPVEGLCLAG GGGLAQQVLV PAGSARPVAF SVVPTAAAAV SLKVVARGSF
0901 EFPVGDAVSK VLQIEKEGAI HREELVYELN PLDHRGRTLE IPGNSDPNMI
0951 PDGDFNSYVR VTASDPLDTL GSEGALSPGG VASLLRLPRG CGEQTMIYLA
1001 PTLAASRYLD KTEQWSTLPP ETKDHAVDLI QKGYMRIQQF RKADGSYAAW
1051 LSRDSSTWLT AFVLKVLSLA QEQVGGSPEK LQETSNWLLS QQQADGSFQD
1101 PCPVLDRSMQ GGLVGNDETV ALTAFVTIAL HHGLAVFQDE GAEPLKQRVE
1151 ASISKANSFL GEKASAGLLG AHAAAITAYA LSLTKAPVDL LGVAHNNLMA
1201 MAQETGDNLY WGSVTGSQSN AVSPTPAPRN PSDPMPQAPA LWIETTAYAL
1251 LHLLLHEGKA EMADQASAWL TRQGSFQGGF RSTQDTVIAL DALSAYWIAS
1301 HTTEERGLNV TLSSTGRNGF KSHALQLNNR QIRGLEEELQ FSLGSKINVK
1351 VGGNSKGTLK VLRTYNVLDM KNTTCQDLQI EVTVKGHVEY TMEANEDYED
1401 YEYDELPAKD DPD APLQPVT PLQLFEGRRN RRRREAPKVV EEQESRVHYT
1451 VCIWRNGKVG LSGMAIADVT LLSGFHALRA DLEKLTSLSD RYVSHFETEG
1501 PHVLLYFDSV PTSRECVGFE AVQEVPVGLV QPASATLYDY YNPERRCSVF
1551 YGAPSKSRLL ATLCSAEVCQ CAEGKCPRQR RALERGLQDE DGYRMKFACY
1601 YPRVEYGFQV KVLREDSRAA FRLFETKITQ VLHFTKDVKA AANQMRNFLV
1651 RASCRLRLEP GKEYLIMGLD GATYDLEGHP QYLLDSNSWI EEMPSERLCR
1701 STRQRAACAQ LNDFLQEYGT QGCQV (SEQ ID NO: 19)

Complement C4-derived peptide CO4-2
APLQPVTPLQ LFEGRRN (SEQ ID NO: 22)

[13] Complement C4-B-derived peptide CO4-2
Peptide CO4-2 of SEQ ID NO: 22 is an amino acid sequence present in the topological region portion common to the proteins of Complement C4-A (SEQ ID NO: 19) and Complement C4-B based on MS / MS and MASCOT database search results. .
Complement C4-B protein is divided into Complement C4 beta chain, Complement C4-B alpha chain, and Complement C4 gamma chain by processing after biosynthesis. SEQ ID NO: 21 is the amino acid sequence of intact protein Complement C4-B, which includes all of these.
Intact protein / peptide
0001 KPRLLLFSPS VVHLGVPLSV GVQLQDVPRG QVVKGSVFLR NPSRNNVPCS
0051 PKVDFTLSSE RDFALLSLQV PLKDAKSCGL HQLLRGPEVQ LVAHSPWLKD
0101 SLSRTTNIQG INLLFSSRRG HLFLQTDQPI YNPGQRVRYR VFALDQKMRP
0151 STDTITVMVE NSHGLRVRKK EVYMPSSIFQ DDFVIPDISE PGTWKISARF
0201 SDGLESNSST QFEVKKYVLP NFEVKITPGK PYILTVPGHL DEMQLDIQAR
0251 YIYGKPVQGV AYVRFGLLDE DGKKTFFRGL ESQTKLVNGQ SHISLSKAEF
0301 QDALEKLNMG ITDLQGLRLY VAAAIIESPG GEMEEAELTS WYFVSSPFSL
0351 DLSKTKRHLV PGAPFLLQAL VREMSGSPAS GIPVKVSATV SSPGSVPEVQ
0401 DIQQNTDGSG QVSIPIIIPQ TISELQLSVS AGSPHPAIAR LTVAAPPSGG
0451 PGFLSIERPD SRPPRVGDTL NLNLRAVGSG ATFSHYYYMI LSRGQIVFMN
0501 REPKRTLTSV SVFVDHHLAP SFYFVAFYYH GDHPVANSLR VDVQAGACEG
0551 KLELSVDGAK QYRNGESVKL HLETDSLALV ALGALDTALY AAGSKSHKPL
0601 NMGKVFEAMN SYDLGCGPGG GDSALQVFQA AGLAFSDGDQ WTLSRKRLSC
0651 PKEKTTRKKR NVNFQKAINE KLGQYASPTA KRCCQDGVTR LPMMRSCEQR
0701 AARVQQPDCR EPFLSCCQFA ESLRKKSRDK GQAGLQRALE ILQEEDLIDE
0751 DDIPVRSFFP ENWLWRVETV DRFQILTLWL PDSLTTWEIH GLSLSKTKGL
0801 CVATPVQLRV FREFHLHLRL PMSVRRFEQL ELRPVLYNYL DKNLTVSVHV
0851 SPVEGLCLAG GGGLAQQVLV PAGSARPVAF SVVPTAAAAV SLKVVARGSF
0901 EFPVGDAVSK VLQIEKEGAI HREELVYELN PLDHRGRTLE IPGNSDPNMI
0951 PDGDFNSYVR VTASDPLDTL GSEGALSPGG VASLLRLPRG CGEQTMIYLA
1001 PTLAASRYLD KTEQWSTLPP ETKDHAVDLI QKGYMRIQQF RKADGSYAAW
1051 LSRDSSTWLT AFVLKVLSLA QEQVGGSPEK LQETSNWLLS QQQADGSFQD
1101 LSPVIHRSMQ GGLVGNDETV ALTAFVTIAL HHGLAVFQDE GAEPLKQRVE
1151 ASISKANSFL GEKASAGLLG AHAAAITAYA LSLTKAPVDL LGVAHNNLMA
1201 MAQETGDNLY WGSVTGSQSN AVSPTPAPRN PSDPMPQAPA LWIETTAYAL
1251 LHLLLHEGKA EMADQASAWL TRQGSFQGGF RSTQDTVIAL DALSAYWIAS
1301 HTTEERGLNV TLSSTGRNGF KSHALQLNNR QIRGLEEELQ FSLGSKINVK
1351 VGGNSKGTLK VLRTYNVLDM KNTTCQDLQI EVTVKGHVEY TMEANEDYED
1401 YEYDELPAKD DPD APLQPVT PLQLFEGRRN RRRREAPKVV EEQESRVHYT
1451 VCIWRNGKVG LSGMAIADVT LLSGFHALRA DLEKLTSLSD RYVSHFETEG
1501 PHVLLYFDSV PTSRECVGFE AVQEVPVGLV QPASATLYDY YNPERRCSVF
1551 YGAPSKSRLL ATLCSAEVCQ CAEGKCPRQR RALERGLQDE DGYRMKFACY
1601 YPRVEYGFQV KVLREDSRAA FRLFETKITQ VLHFTKDVKA AANQMRNFLV
1651 RASCRLRLEP GKEYLIMGLD GATYDLEGHP QYLLDSNSWI EEMPSERLCR
1701 STRQRAACAQ LNDFLQEYGT QGCQV (SEQ ID NO: 21)

As a precaution, the sequence of CO4-2 is shown below.
Complement C4-derived peptide CO4-2
APLQPVTPLQ LFEGRRN (SEQ ID NO: 22)

[14] Fibrinogen alpha chain (isoform 1) derived peptide FIBA-1
The area value of FIBA-1 of SEQ ID NO: 24 is statistically significantly (p <0.05) higher in cognitive impairment disorders (AD, MCI, DLB, FTD) as a result of t-test than NDC (FIG. 15), FIBA-1 of SEQ ID NO: 24 may be useful in distinguishing between patients with cognitive impairment disorders (AD, MCI, DLB, FTD) and subjects who do not suffer from mental disorders (NDC) I understood. Analysis by recipient operating characteristic curves clearly proves useful in distinguishing AD and MCI patients from subjects not suffering from mental illness (Table 1).
Intact protein / peptide
0001 GPRVVERHQS ACKDSDWPFC SDEDWNYKCP SGCRMKGLID EVNQDFTNRI
0051 NKLKNSLFEY QKNNKDSHSL TTNIMEILRG DFSSANNRDN TYNRVSEDLR
0101 SRIEVLKRKV IEKVQHIQLL QKNVRAQLVD MKRLEVDIDI KIRSCRGSCS
0151 RALAREVDLK DYEDQQKQLE QVIAKDLLPS RDRQHLPLIK MKPVPDLVPG
0201 NFKSQLQKVP PEWKALTDMP QMRMELERPG GNEITRGGST SYGTGSETES
0251 PRNPSSAGSW NSGSSGPGST GNRNPGSSGT GGTATWKPGS SGPGSTGSWN
0301 SGSSGTGSTG NQNPGSPRPG STGTWNPGSS ERGSAGHWTS ESSVSGSTGQ
0351 WHSESGSFRP DSPGSGNARP NNPDWGTFEE VSGNVSPGTR REYHTEKLVT
0401 SKGDKELRTG KEKVTSGSTT TTRRSCSKTV TKTVIGPDGH KEVTKEVVTS
0451 EDGSDCPEAM DLGTLSGIGT LDGFRHRHPD EAAFFDTAST GKTFPGFFSP
0501 MLGEFVSETE SRGSESGIFT NTKESSSHHP GIAEFPSRGK SSSYSKQFTS
0551 STSYNRGDST FES KSYKMAD EAGSEADHEG THSTKRGHAK SRPVRDCDDV
0601 LQTHPSGTQS GIFNIKLPGS SKIFSVYCDQ ETSLGGWLLI QQRMDGSLNF
0651 NRTWQDYKRG FGSLNDEGEG EFWLGNDYLH LLTQRGSVLR VELEDWAGNE
0701 AYAEYHFRVG SEAEGYALQV SSYEGTAGDA LIEGSVEEGA EYTSHNNMQF
0751 STFDRDADQW EENCAEVYGG GWWYNNCQAA NLNGIYYPGG SYDPRNNSPY
0801 EIENGVVWVS FRGADYSLRA VRMKIRPLVT Q (SEQ ID NO: 23)

Fibrinogen alpha chain-derived peptide FIBA-1
SSSYSKQFTS STSYNRGDST FES (SEQ ID NO: 24)

[15] Fibrinogen alpha chain (isoform 2) derived peptide FIBA-1
From the results of MS / MS and MASCOT database search, peptide FIBA-1 of SEQ ID NO: 24 is a topological region portion common to the proteins of Fibrinogen alpha chain (isoform 1) (SEQ ID NO: 23) and Fibrinogen alpha chain (isoform 2) Is the amino acid sequence present in The amino acid sequence of the fibrinogen alpha chain (isoform 2) intact protein is shown below as SEQ ID NO: 25.
Intact protein / peptide
0001 GPRVVERHQS ACKDSDWPFC SDEDWNYKCP SGCRMKGLID EVNQDFTNRI
0051 NKLKNSLFEY QKNNKDSHSL TTNIMEILRG DFSSANNRDN TYNRVSEDLR
0101 SRIEVLKRKV IEKVQHIQLL QKNVRAQLVD MKRLEVDIDI KIRSCRGSCS
0151 RALAREVDLK DYEDQQKQLE QVIAKDLLPS RDRQHLPLIK MKPVPDLVPG
0201 NFKSQLQKVP PEWKALTDMP QMRMELERPG GNEITRGGST SYGTGSETES
0251 PRNPSSAGSW NSGSSGPGST GNRNPGSSGT GGTATWKPGS SGPGSTGSWN
0301 SGSSGTGSTG NQNPGSPRPG STGTWNPGSS ERGSAGHWTS ESSVSGSTGQ
0351 WHSESGSFRP DSPGSGNARP NNPDWGTFEE VSGNVSPGTR REYHTEKLVT
0401 SKGDKELRTG KEKVTSGSTT TTRRSCSKTV TKTVIGPDGH KEVTKEVVTS
0451 EDGSDCPEAM DLGTLSGIGT LDGFRHRHPD EAAFFDTAST GKTFPGFFSP
0501 MLGEFVSETE SRGSESGIFT NTKESSSHHP GIAEFPSRGK SSSYSKQFTS
0551 STSYNRGDST FES KSYKMAD EAGSEADHEG THSTKRGHAK SRPVRGIHTS
0601 PLGKPSLSP (SEQ ID NO: 25)

As a precaution, the sequence of FIBA-1 is shown below.
Fibrinogen alpha chain-derived peptide FIBA-1
SSSYSKQFTS STSYNRGDST FES (SEQ ID NO: 24)

[16] Fibrinogen alpha chain (isoform 1) derived peptide FIBA-2
The area value of FIBA-2 of SEQ ID NO: 26 is statistically significantly (p <0.05) higher in cognitive impairment disorders (AD, MCI, DLB, FTD) as a result of t-test than NDC (FIG. 16), FIBA-2 of SEQ ID NO: 26 is useful in distinguishing between patients with cognitive dysfunction disorders (AD, MCI, DLB, FTD) and subjects without mental illness (NDC) I understood. Analysis by recipient operating characteristic curves clearly proves useful in distinguishing AD and MCI patients from subjects not suffering from mental illness (Table 1).
Intact protein / peptide
0001 GPRVVERHQS ACKDSDWPFC SDEDWNYKCP SGCRMKGLID EVNQDFTNRI
0051 NKLKNSLFEY QKNNKDSHSL TTNIMEILRG DFSSANNRDN TYNRVSEDLR
0101 SRIEVLKRKV IEKVQHIQLL QKNVRAQLVD MKRLEVDIDI KIRSCRGSCS
0151 RALAREVDLK DYEDQQKQLE QVIAKDLLPS RDRQHLPLIK MKPVPDLVPG
0201 NFKSQLQKVP PEWKALTDMP QMRMELERPG GNEITRGGST SYGTGSETES
0251 PRNPSSAGSW NSGSSGPGST GNRNPGSSGT GGTATWKPGS SGPGSTGSWN
0301 SGSSGTGSTG NQNPGSPRPG STGTWNPGSS ERGSAGHWTS ESSVSGSTGQ
0351 WHSESGSFRP DSPGSGNARP NNPDWGTFEE VSGNVSPGTR REYHTEKLVT
0401 SKGDKELRTG KEKVTSGSTT TTRRSCSKTV TKTVIGPDGH KEVTKEVVTS
0451 EDGSDCPEAM DLGTLSGIGT LDGFRHRHPD EAAFFDTAST GKTFPGFFSP
0501 MLGEFVSETE SRGSESGIFT NTKESSSHHP GIAEFPSRGK SSSYSKQFTS
0551 STSYNRGDST FESKS YKMAD EAGSEADHEG THSTKRGHAK SRPVRDCDDV
0601 LQTHPSGTQS GIFNIKLPGS SKIFSVYCDQ ETSLGGWLLI QQRMDGSLNF
0651 NRTWQDYKRG FGSLNDEGEG EFWLGNDYLH LLTQRGSVLR VELEDWAGNE
0701 AYAEYHFRVG SEAEGYALQV SSYEGTAGDA LIEGSVEEGA EYTSHNNMQF
0751 STFDRDADQW EENCAEVYGG GWWYNNCQAA NLNGIYYPGG SYDPRNNSPY
0801 EIENGVVWVS FRGADYSLRA VRMKIRPLVT Q (SEQ ID NO: 23)

Fibrinogen alpha chain derived peptide FIBA-2
SSSYSKQFTS STSYNRGDST FESKS (SEQ ID NO: 26)

[17] Fibrinogen alpha chain (isoform 2) derived peptide FIBA-2
Peptide FIBA-2 of SEQ ID NO: 26 is a topological region part common to proteins of Fibrinogen alpha chain (isoform 1) (SEQ ID NO: 23) and Fibrinogen alpha chain (isoform 2) based on MS / MS and MASCOT database search results Is the amino acid sequence present in The amino acid sequence of the fibrinogen alpha chain (isoform 2) intact protein is shown below as SEQ ID NO: 25.
Intact protein / peptide
0001 GPRVVERHQS ACKDSDWPFC SDEDWNYKCP SGCRMKGLID EVNQDFTNRI
0051 NKLKNSLFEY QKNNKDSHSL TTNIMEILRG DFSSANNRDN TYNRVSEDLR
0101 SRIEVLKRKV IEKVQHIQLL QKNVRAQLVD MKRLEVDIDI KIRSCRGSCS
0151 RALAREVDLK DYEDQQKQLE QVIAKDLLPS RDRQHLPLIK MKPVPDLVPG
0201 NFKSQLQKVP PEWKALTDMP QMRMELERPG GNEITRGGST SYGTGSETES
0251 PRNPSSAGSW NSGSSGPGST GNRNPGSSGT GGTATWKPGS SGPGSTGSWN
0301 SGSSGTGSTG NQNPGSPRPG STGTWNPGSS ERGSAGHWTS ESSVSGSTGQ
0351 WHSESGSFRP DSPGSGNARP NNPDWGTFEE VSGNVSPGTR REYHTEKLVT
0401 SKGDKELRTG KEKVTSGSTT TTRRSCSKTV TKTVIGPDGH KEVTKEVVTS
0451 EDGSDCPEAM DLGTLSGIGT LDGFRHRHPD EAAFFDTAST GKTFPGFFSP
0501 MLGEFVSETE SRGSESGIFT NTKESSSHHP GIAEFPSRGK SSSYSKQFTS
0551 STSYNRGDST FESKS YKMAD EAGSEADHEG THSTKRGHAK SRPVRGIHTS
0601 PLGKPSLSP (SEQ ID NO: 25)

As a precaution, the sequence of FIBA-2 is shown below.
Fibrinogen alpha chain derived peptide FIBA-2
SSSYSKQFTS STSYNRGDST FESKS (SEQ ID NO: 26)

[18] Fibrinogen alpha chain (isoform 1) derived peptide FIBA-3
The area value of FIBA-3 of SEQ ID NO: 27 is statistically significantly (p <0.05) higher in cognitive dysfunction diseases (AD, MCI, DLB, FTD) than in NDC as a result of t-test. (FIG. 17), FIBA-3 of SEQ ID NO: 27 is useful in distinguishing between cognitive dysfunction disease (AD, MCI, DLB, FTD) patients and subjects not suffering from mental disorders (NDC) I understood. Analysis by recipient operating characteristic curves clearly proves useful in distinguishing AD and MCI patients from subjects not suffering from mental illness (Table 1).
Intact protein / peptide
0001 GPRVVERHQS ACKDSDWPFC SDEDWNYKCP SGCRMKGLID EVNQDFTNRI
0051 NKLKNSLFEY QKNNKDSHSL TTNIMEILRG DFSSANNRDN TYNRVSEDLR
0101 SRIEVLKRKV IEKVQHIQLL QKNVRAQLVD MKRLEVDIDI KIRSCRGSCS
0151 RALAREVDLK DYEDQQKQLE QVIAKDLLPS RDRQHLPLIK MKPVPDLVPG
0201 NFKSQLQKVP PEWKALTDMP QMRMELERPG GNEITRGGST SYGTGSETES
0251 PRNPSSAGSW NSGSSGPGST GNRNPGSSGT GGTATWKPGS SGPGSTGSWN
0301 SGSSGTGSTG NQNPGSPRPG STGTWNPGSS ERGSAGHWTS ESSVSGSTGQ
0351 WHSESGSFRP DSPGSGNARP NNPDWGTFEE VSGNVSPGTR REYHTEKLVT
0401 SKGDKELRTG KEKVTSGSTT TTRRSCSKTV TKTVIGPDGH KEVTKEVVTS
0451 EDGSDCPEAM DLGTLSGIGT LDGFRHRHPD EAAFFDTAST GKTFPGFFSP
0501 MLGEFVSETE SRGSESGIFT NTKESSSHHP GIAEFPSRGK SSSYSKQFTS
0551 STSYNRGDST FESKSY KMAD EAGSEADHEG THSTKRGHAK SRPVRDCDDV
0601 LQTHPSGTQS GIFNIKLPGS SKIFSVYCDQ ETSLGGWLLI QQRMDGSLNF
0651 NRTWQDYKRG FGSLNDEGEG EFWLGNDYLH LLTQRGSVLR VELEDWAGNE
0701 AYAEYHFRVG SEAEGYALQV SSYEGTAGDA LIEGSVEEGA EYTSHNNMQF
0751 STFDRDADQW EENCAEVYGG GWWYNNCQAA NLNGIYYPGG SYDPRNNSPY
0801 EIENGVVWVS FRGADYSLRA VRMKIRPLVT Q (SEQ ID NO: 23)

Fibrinogen alpha chain-derived peptide FIBA-3
SSSYSKQFTS STSYNRGDST FESKSY (SEQ ID NO: 27)

[19] Fibrinogen alpha chain (isoform 2) derived peptide FIBA-3
The peptide FIBA-3 of SEQ ID NO: 26 is a topological region part common to the proteins of Fibrinogen alpha chain (isoform 1) (SEQ ID NO: 23) and Fibrinogen alpha chain (isoform 2) based on the results of MS / MS and MASCOT database searches. Is the amino acid sequence present in The amino acid sequence of the fibrinogen alpha chain (isoform 2) intact protein is shown below as SEQ ID NO: 25.
Intact protein / peptide
0001 GPRVVERHQS ACKDSDWPFC SDEDWNYKCP SGCRMKGLID EVNQDFTNRI
0051 NKLKNSLFEY QKNNKDSHSL TTNIMEILRG DFSSANNRDN TYNRVSEDLR
0101 SRIEVLKRKV IEKVQHIQLL QKNVRAQLVD MKRLEVDIDI KIRSCRGSCS
0151 RALAREVDLK DYEDQQKQLE QVIAKDLLPS RDRQHLPLIK MKPVPDLVPG
0201 NFKSQLQKVP PEWKALTDMP QMRMELERPG GNEITRGGST SYGTGSETES
0251 PRNPSSAGSW NSGSSGPGST GNRNPGSSGT GGTATWKPGS SGPGSTGSWN
0301 SGSSGTGSTG NQNPGSPRPG STGTWNPGSS ERGSAGHWTS ESSVSGSTGQ
0351 WHSESGSFRP DSPGSGNARP NNPDWGTFEE VSGNVSPGTR REYHTEKLVT
0401 SKGDKELRTG KEKVTSGSTT TTRRSCSKTV TKTVIGPDGH KEVTKEVVTS
0451 EDGSDCPEAM DLGTLSGIGT LDGFRHRHPD EAAFFDTAST GKTFPGFFSP
0501 MLGEFVSETE SRGSESGIFT NTKESSSHHP GIAEFPSRGK SSSYSKQFTS
0551 STSYNRGDST FESKSY KMAD EAGSEADHEG THSTKRGHAK SRPVRGIHTS
0601 PLGKPSLSP (SEQ ID NO: 25)

As a precaution, the sequence of FIBA-3 is shown below.
Fibrinogen alpha chain-derived peptide FIBA-3
SSSYSKQFTS STSYNRGDST FESKSY (SEQ ID NO: 27)

  Table 1 shows the AUC values obtained by the analysis using the receiver operating characteristic curve in the detection of cognitive impairment disease of each marker peptide.

  Mass spectrometry or immunology of serum or plasma abundance directly, with or without liquid chromatography and / or other suitable separation means, using each of these marker peptides alone or in combination It is possible to distinguish cognitive dysfunction diseases such as AD, MCI, DLB, and FTD from non-psychiatric subjects including healthy individuals by measuring by other methods such as mechanical or enzymological methods. is there.

  Since the biomarker disclosed in the present invention can be used to detect cognitive impairment disorders including mild cognitive impairment and Alzheimer's disease, it can be applied to applications in the diagnostic field including diagnostic agents.

Claims (7)

  A biomarker for diagnosis of cognitive dysfunction disease, which is a transcription factor AP-2 gamma-derived peptide AP2C consisting of the amino acid sequence represented by SEQ ID NO: 4.   A peptide comprising the amino acid sequence represented by SEQ ID NO: 4, which appears or increases in a biological sample of a patient not suffering from mental illness in a biological sample of a cognitive dysfunction disease patient marker.   An Alzheimer's disease biomarker that is a peptide consisting of the amino acid sequence represented by SEQ ID NO: 4 and that appears or increases in a biological sample of an Alzheimer's disease patient as compared to a biological sample of a subject not suffering from a mental illness.   A mild cognitive impairment biomarker that is a peptide comprising the amino acid sequence represented by SEQ ID NO: 4 and that appears or increases in a biological sample of a patient with mild cognitive impairment as compared to a biological sample of a subject not suffering from mental illness.   A method for detecting a biomarker for cognitive dysfunction disease diagnosis, comprising measuring the biomarker for cognitive dysfunction disease diagnosis according to any one of claims 1 to 4 in a biological sample.   The bio for cognitive dysfunction disease according to claim 5, wherein the detection is performed by immunoblotting or Western blotting, enzyme or fluorescent or radioactive substance-labeled antibody method, mass spectrometry, immunoMS method or surface plasmon resonance method. Marker detection method.   A detection kit for a cognitive dysfunction disease comprising an antibody or aptamer to the biomarker according to any one of claims 1 to 4.