TWI606060B - Peptides for detecting anti-α7 nachr antibody - Google Patents

Description

Polypeptide molecule for detecting alpha seven-unit antibody of neurothyroid acetylcholine receptor

The present invention relates to a polypeptide molecule for detecting an antibody, and more particularly to a polypeptide molecule for detecting a neuronal acetylcholine receptor subunit alpha-7 ( α 7 nAChR) antibody.

With the changes of society, the pace of life is becoming more and more compact, and the pressure is accompanied, which makes the incidence of various Psychiatric disorders increase. Among them, Schizophrenia is a relatively common and serious mental illness with a lifetime prevalence of 1%.

Over the years, doctors have obtained information such as medical history and mental state through clinical observation and interviews, and then judged whether individuals suffer from schizophrenia based on symptomology. However, this type of diagnosis is greatly influenced by subjective factors. Inevitably, misdiagnosis and missed diagnosis often occur, which is one of the important reasons for the current criticism of schizophrenia disease diagnosis methods. Therefore, an objective detection index for schizophrenia is urgently needed as a supplementary basis for clinicians to comprehensively evaluate whether or not they suffer from schizophrenia.

In view of the defects of the prior art, the present invention provides a linear polypeptide molecule of a simple 7-unit unit epitope of a neurogenic acetylcholine receptor, which can be combined with a neurogenic acetylcholine receptor alpha 7-unit antibody. To detect the content of alpha 7-unit antibody (ie, autoantibody) in the blood, plasma, or serum of an individual, and to diagnose early schizophrenia.

In addition, the present invention provides a neurogenic acetylcholine receptor alpha 7-unit polypeptide molecule which is low in cost and easy to preserve, and the prepared detection reagent and immunoenzyme assay kit are easy to store.

The invention also provides a reagent and an immunoenzyme assay kit for facilitating detection of schizophrenia, thereby increasing the convenience and rapidity of use.

In a preferred embodiment, the present invention provides an antibody binding molecule which binds to an antibody of a neuronal acetylcholine receptor subunit alpha-7; wherein the antibody binds The molecule is 25-40 amino acids in length, and the sequence of the antibody binding molecule comprises at least one of SEQ ID NOS: 1 to 5.

In a preferred embodiment, the antibody binding molecule has a sequence length of from 25 to 35 amino acids.

In a preferred embodiment, the amount of proline in the antibody binding molecule sequence is from 0 to 3.

In a preferred embodiment, the amount of glutamic acid (Glutamic acid) in the antibody binding molecule sequence is 0 to 3, or the number of Cysteine in the sequence. It is 0 to 1.

In a preferred embodiment, the antibody binds to a linear polypeptide molecule of the molecule, or the antibody binding molecule does not form an alpha helix structure in an aqueous solution or a plasma.

In a preferred embodiment, the binding strength of the antibody binding molecule to the antibody is greater than the interaction between the antibody and a wash buffer.

The present invention further provides a detection reagent comprising an antibody binding molecule having a length of 25-40 amino acids, and the sequence of the antibody binding molecule comprises at least one of SEQ ID NOS: 1 to 5.

The invention further provides an immunoassay assay kit (ELISA Kit) comprising a reagent; wherein the reagent further comprises an antibody binding molecule, the antibody binding molecule has a length of 25-40 amino acids, and the antibody binds The sequence of the molecule comprises at least one of SEQ ID NOS: 1 to 5.

In a preferred embodiment, the concentration of the antibody binding molecule in the reagent is from 0.75 μg/mL to 5 mg/mL.

The invention further provides an immunological enzyme assay kit (ELISA Kit) comprising a porous disk; wherein the porous disk comprises an antibody binding molecule having a length of 25-40 amino acids, and the antibody binding molecule The sequence of the antibody binding molecule comprises at least one of SEQ ID NOS: 1 to 5.

In a preferred embodiment, the immunoenzyme assay kit is used to detect the antibody content of an alpha 7-unit unit of a neurotransmitter in a blood, a plasma, or a serum.

In a preferred embodiment, the immunoenzyme assay kit can be used for early diagnosis of a mental illness or as an indicator of a condition of a mental illness.

In a preferred embodiment, the mental illness is schizophrenia.

Figure 1: Receiver operating characteristic curves plotted against the SBI values of the schizophrenia group and its matching healthy group.

Schizophrenia is a neuropsychiatric disorder. Although the cause of the disease is still unclear, it is known that schizophrenia is closely related to individual immune dysfunction, autoimmune response and inflammatory response (Strous and Shoenfeld, 2006; Potvin et al., 2008). Moreover, the cellular immune function of patients with schizophrenia is decreased, and the humoral immune function is significantly increased (Potvin et al., 2008).

Alpha-7 nicotinic receptor (α7 receptor) is an ion channel type receptor with high selectivity for calcium ions. (Neuronal acetylcholine receptor subunit alpha-7, abbreviated as α 7 nAChR, which is a protein end product expressed after CHRNA7 gene expression), expressed in the cerebral cortex, hippocampus (Hippocampus; Hellstrom-Lindahl et al., 1999), and lymphocytes Cells (Lymphocyte; Wang et al., 2001; Villiger et al., 2002) play an important role in signal transmission, learning, memory, synaptic plasticity, and immune response between neurons.

Studies have shown that the loss of the neuroic acetylcholine receptor alpha 7-unit gene (CHRNA7 gene) is closely related to schizophrenia (Shinawi et al., 2009). In addition, 23% more schizophrenic patient neuropathic acetylcholine alpha 7 receptors subunits (α 7 nAChR) autoantibodies (autoantibody) was significantly higher than the average person (Chandley et al., 2009) , may be hindered Nicotine receptor function is broken, which in turn is involved in the pathogenesis of schizophrenia. Based on the research results and experience of the inventors for many years, it is believed that a simple and accurate test method should be established to detect schizophrenia disease early by detecting the content of α 7 nAChR autoantibodies in the blood.

The following is a detailed description of embodiments of the invention, as well as the techniques and features of the invention. The present invention is not intended to limit the invention, and any changes and modifications made by those skilled in the art without departing from the spirit and scope of the invention are included in the scope of the invention.

The present invention provides at least one antibody binding molecule, which can be combined with an antibody of Neuronal acetylcholine receptor subunit alpha-7 ( α 7 nAChR) to detect blood, plasma, or The serum acetylcholine in the serum is subjected to an alpha 7-unit antibody (ie, autoantibody), which in turn diagnoses schizophrenia or serves as an indicator for tracking schizophrenia.

The antibody binding molecule of the present invention may be a neurogenic acetylcholine receptor alpha 7 nARR Recombinant, a neurogenic acetylcholine receptor alpha 7 subunit epitope (Epitope) polypeptide molecule, or a nerve A acetylcholine receptor alpha alpha epitope linear polypeptide molecule. Among them, compared with the neurogenic acetylcholine receptor, the 7-unit recombinant protein needs to be constructed, transfected, expressed, screened, purified, etc., and the spatial structure of the recombinant protein is complex, preferably, neurotic. Acetylcholine receptor alpha subunit 7 epitope linear polypeptide molecule (hereinafter part will be referred to as α 7 nAChR epitope linear polypeptide molecule).

Since antigenic antibodies are complementary in spatial and spatial configuration, the combination of the two actually occurs only between the epitope (Epitope) and the antibody binding site (Paratope). Therefore, only the epitope fragment can represent the state and affinity of the entire protein to bind to the antibody. According to the inventor's many years of experience and research results, it is believed that the 7-unit epitope of the neurogenic acetylcholine receptor can be found from the sequence and structure of the 7-unit unit of the neuroacetylcholine receptor. Screening appropriate " α 7 nAChR epitope linear polypeptide molecules" from a variety of neurogenic acetylcholine receptor alpha 7-unit epitope sequences to develop rapid, effective, and stable schizophrenia detection reagents and schizophrenia Immunoassay kit (ELISA Kit).

Neuroic acetylcholine receptor alpha 7-unit epitope linear polypeptide molecule design:

The inventors used bioinformatics to analyze the amino acid sequence of the 7-unit unit of the neuroacetylcholine receptor, and then predicted the antigen-related parameters by multiple epitope prediction software, and then selected the candidate sequences. Preferably, the candidate sequences are 25 to 40 amino acids in length, or, preferably, the candidate sequences are 25 to 35 amino acids in length.

The immunoenzymatic assay (ELISA) is highly sensitive and therefore requires a high degree of stability to the amino acid sequence molecule. The inventors further followed the following seven principles to screen candidate sequences:

1) Select a sequence that does not form an alpha helix structure. Preferably, the alpha helix structure is not formed in an aqueous solution or a plasma.

2) The peptides at both ends are arranged better than the middle.

3) Avoid internal repeats of the protein.

4) Avoid peptides with strong homology.

5) Avoid cysteine (Cysteine) and glutamic acid (Glutamic acid) in the sequence. Preferably, the number of glutamic acid in the sequence is from 0 to 3, or, preferably, the number of Cysteine in the sequence is from 0 to 1.

6) Avoid excessive proline (Proline), but having 1-2 proline in the sequence facilitates the stability of the peptide chain structure and is beneficial for the production of specific antibodies. Preferably, the number of proline in the sequence is from 0 to 3.

7) Preferably, it must contain a restriction epitope of the Human leukocyte antigen class II (HLA class II) system, including HLA-DR, HLA-DP, and HLA-DQ. gauge. These epitopes can be recognized by the second type human leukocyte antigen system of more than 90% of the Chinese population.

The inventors finally screened the following five neurogenic acetylcholine receptor alpha 7-element epitope linear polymorphisms, which are SEQ ID NOs: 1 to 5 in the sequence listing:

SEQ ID NO 1: NH ₃ ⁺ - LTVYFSLSLLQIMDVLTTNIWLQMSWTDHYLQK -COO ^-

SEQ ID NO 2: NH ₃ ⁺ - VSLQGEFQRKLYKELVKNYNPLERPVANDSQPL -COO ^-

SEQ ID NO 3: NH ₃ ⁺ - EMPKWTRVILLNWCAWFLRMKRPGED -COO ^-

SEQ ID NO 4: NH ₃ ⁺ - RLMAFSVFTIICTIGILMSAPNFVEAVSK -COO ^-

SEQ ID NO 5: NH ₃ ⁺ - KNQVLTTNIWLQMSWTDHYLQWNVSCR -COO ^-

Preparation of a linear acetylcholine receptor alpha 7-unit epitope linear polypeptide molecule:

The neuropeptides of the alpha acetylcholine receptor are linearly synthesized by a chemical method. Preferably, the purity must be greater than 95%. Preferably, the pH is greater than 7. The peptide synthesis synthesis service company can be commissioned to synthesize the linear peptide molecules of the α 7 nAChR epitope, for example, entrusted Mingxin Biotechnology Co., Ltd. (Address: 1st Floor, 10th Floor, No. 3, Park Street, Nangang District, Taipei; Tel: 02-26557128; Http://www.missionbio.com.tw/) Preparation.

The synthetic neuroacetylcholine receptor alpha 7-element epitope linear polypeptide molecule was dissolved in 67% acetic acid (5 mg α 7 nAChR epitope linear polypeptide molecule / 1 mL of 67% acetic acid) at -20 ° C Store in a low temperature freezer.

Neuropeptide acetylcholine receptor alpha 7-unit epitope linear peptide molecular binding analysis:

The binding of the linear 7-element epitope linear polypeptide molecule to the neuroacetylcholine receptor alpha 7-unit antibody was performed by immuno-enzyme assay (ELISA).

First, various coating liquid concentrations were adjusted to 7.5 to 12.5 μg/mL in 0.01 M PBS (pH 7.0 to 7.4). 0.01 M PBS in 1 liter of deionized water contained 0.2964 g of NaH ₂ PO ₄ ‧2H ₂ O, 2.9 g of Na ₂ HPO ₄ ‧12H ₂ O, 8 g of NaCl. Preferably, the 0.01 M PBS contains 0.1% NaN ₃ .

Wherein, the experimental antigen coating solution 1 comprises the α 7 nAChR epitope linear polypeptide molecule of SEQ ID NO: 1 of the present invention. The experimental antigen coating solution 2 is the α 7 nAChR epitope linear polypeptide molecule of SEQ ID NO: 2 of the present invention. The experimental antigen coating solution 3 is the α 7 nAChR epitope linear polypeptide molecule of SEQ ID NO: 3 of the present invention. The experimental antigen coating solution 4 is the α 7 nAChR epitope linear polypeptide molecule of SEQ ID NO: 4 of the present invention. The experimental antigen coating solution 5 is the α 7 nAChR epitope linear polypeptide molecule of SEQ ID NO: 5 of the present invention. Positive control antigen coating human neurogenic acetylcholine receptor alpha 7-unit recombinant protein (Cat. No. H00001139-P01, Abnova). Negative control antigen coating solution The linear polypeptide molecule designed by the inventor has no homology with the human neurogenic acetylcholine receptor alpha 7-unit, so it will not be associated with the acetylcholine receptor alpha 7 unit. Antibody cross-binding.

Each of the above-mentioned various coating liquids was coated on a porous disk, 100 μL per well, and allowed to stand overnight at 4 ° C (Overnight). Preferably, the porous disk is a 96-well disk, but is not limited thereto. Taking the 96-well plate (Cat. No. 224-0096, COSTAR) as an example, the experimental antigen coating solution 1 was coated in two wells of column A, which was the experimental antigen group 1. The experimental antigen coating solution 2 was coated in two wells of column B, which was the experimental antigen group 2. The experimental antigen coating solution 3 was coated in two wells of the C column, which was the experimental antigen group 3. The experimental antigen coating solution 4 was coated in two wells of column D, which was the experimental antigen group 4. The experimental antigen coating solution 5 was coated in two wells of the E column, which was the experimental antigen group 5. A positive control antigen coating solution was coated in two wells of column F, which was a positive control antigen group. A negative control antigen coating solution was coated in two wells of column G, which was a negative control antigen group. 0.01 M PBS (pH 7.0 to 7.4) was coated in two wells of column H, which was a negative control group.

Wash 3 times with 0.01 M PBS (additionally 0.05% TWEEN-20). That is, it was washed 3 times with a washing buffer.

Dilute human neurotransmitter acetylcholine receptor alpha 7-unit antibody (LifeSpan BioSciences) using sample dilution assay (1% BSA in 0.01 M PBS) 100 to 500 times, 100 μL per well, and culture at 25 ° C for 2 to 3 hours.

Wash 3 times with 0.01 M PBS (additionally 0.005% TWEEN-20). That is, it was washed 3 times with a washing buffer. Wherein, the binding ability of the linear polypeptide molecule of the CHRNA7 epitope to the antibody of the neurogenic acetylcholine receptor alpha 7-unit multi-strain is greater than the neurogenic acetylcholine receptor alpha 7-unit multi-drug antibody and the washing buffer The interaction between the two, so the neurogenic acetylcholine receptor alpha 7 units of multiple antibodies will not be washed away by the wash buffer.

200 μL of goat anti-rabbit IgG antibody (Sigma) with horseradish peroxidase (HRP) was added and cultured at 25 ° C for 2 hours.

After washing with 0.01 M PBS, 100 μL of 3,3',5,5'-tetramethylbenzidine (TMB; Invtrogen) was added, and the temperature was kept at room temperature for 15 to 30 minutes.

The reaction was terminated by the addition of 50 μL of 10% H ₂ SO ₄ and the absorbance (OD value) was measured. The detection wavelength is 450 nm, the reference wavelength is 630 nm, and the detection must be completed within 10 minutes.

After subtracting the absorbance values of the negative control groups from each group, the experimental antigen groups 1, 2, 3, 4, 5 and the negative control antigen groups were analyzed for significant difference (p<0.01), and the positive control antigen group and There was a significant difference between the negative control antigen groups (p < 0.001).

Neuroacridine choline receptor alpha 7-unit epitope linear peptide molecule detection of schizophrenia effect analysis:

A, sample collection:

Plasma samples from 306 patients with schizophrenia and 547 healthy human serum samples with age and gender were matched. Clinical diagnosis of schizophrenia in line with the American Psychiatric Association DSM-IV diagnostic criteria.

B. Detection method:

The autoimmune assay (ELISA) was used to detect the autoantibody content of patients with schizophrenia. The method and method for detecting autoantibodies in a schizophrenia patient are substantially the same as the method for synthesizing a linear polypeptide molecule of an alpha 7-unit epitope in the above neurocholine acetylcholine receptor. The steps are as follows: First, the SEQ ID NO: 1, 2, 3, 4, 5 neuroacetylcholine receptors alpha 7 times unit epitope linear polypeptide molecules are dissolved, and mixed in equal proportions. The concentration of the linear acetylcholine receptor alpha-subunit epitope linear polypeptide molecule mixture was adjusted to 7.5-15 μg/mL in 0.01 M PBS (pH 7.0-7.4). Preferably, the 0.01 M PBS contains 0.1% NaN ₃ .

The concentrations of the positive control antigen and the negative control antigen were adjusted to 15-20 μg/mL in 0.01 M PBS (pH 7.0 to 7.4), respectively. Preferably, the 0.01 M PBS contains 0.1% NaN ₃ .

Each of the coating liquids was individually coated on a porous disk, 100 μL per well, and allowed to stand overnight at 4 ° C (Overnight). Preferably, the porous disk is a 96-well disk, but is not limited thereto. Taking a 96-well plate (Cat. No. 224-0096, COSTAR) as an example, a mixture of a linear acetylcholine receptor alpha-subunit epitope linear polypeptide molecule is coated in two wells of column A, Experimental antigen group. A positive control antigen coating solution was coated in two wells of column B, which was a positive control antigen group. The negative control antigen coating solution was coated in two wells of column C, which was a negative control antigen group. 0.01 M PBS (pH 7.0 to 7.4) was coated in 2 wells of column D, which was a negative control group.

Wash 3 times with 0.01 M PBS (additionally 0.05% TWEEN-20).

Use the sample dilution assay solution (1% BSA in 0.01M PBS) to apply the spirit The plasma of patients with schizophrenia is diluted 100-500 times, 100 μL per well, and cultured at 25 ° C for 2 to 3 hours.

Wash 3 times with 0.01 M PBS (additionally 0.005% TWEEN-20). 200 μL of goat anti-human IgG antibody (Sigma) with horseradish peroxidase (HRP) was added and incubated at 25 ° C for 2 hours.

After washing with 0.01 M PBS, 100 μL of 3,3',5,5'-tetramethylbenzidine (TMB; Invtrogen) was added, and the temperature was kept at room temperature for 15 to 30 minutes.

The reaction was terminated by the addition of 50 μL of 10% H ₂ SO ₄ and the absorbance (OD value) was measured. The detection wavelength was 450 nm and the reference wavelength was 630 nm.

C, data analysis method:

The specific binding index (SBI) was used to determine the degree of binding of the linear 7-element epitope linear polypeptide molecule to plasma autoantibodies in the neuroacetylcholine receptor.

SBI = [experimental antigen group absorbance - negative control absorbance] / [negative control antigen group absorbance - negative control absorbance].

A Z-test was used to analyze whether there was a difference between the SBI value of plasma samples from patients with schizophrenia and the SBI values of healthy human plasma samples.

D. Experimental results:

Please refer to Table 1. The alpha 7-unit IgG antibody in the plasma of patients with schizophrenia was significantly higher than that of the healthy group, Z=6.27, with statistical difference (p<0.0001).

The above results fully show that the use of this case neuroic acetylcholine receptor alpha 7 The self-antibody content of the linear polypeptide molecules screened by the subunit epitope has significant statistical differences between patients with schizophrenia and healthy people. Therefore, the neuroic acetylcholine receptor alpha 7-element epitope linear polypeptide molecule can be used as an auxiliary tool for clinical detection of schizophrenia to provide autoantibody data as an objective diagnostic indicator.

Analysis of the accuracy of linear peptide detection of alpha 7-unit epitopes in neurogenic acetylcholine receptors:

A. Detection method: The Receiver operating characteristic curve (ROC) was used to evaluate the linear peptide molecular detection effect of the 7-unit epitope of the neuroacetylcholine receptor in this case. The receiver operating characteristic curve of the analysis is to set different thresholds sequentially, and the detection result of the individual whose SBI value is higher than the threshold value is defined as positive, and the detection result of the individual whose SBI value is lower than the threshold value is defined as negative.

Among them, those with schizophrenia test results are positive, further defined as true positive. Patients with schizophrenia whose test results are negative are further defined as pseudo-negative.

Those whose health test results are positive are further defined as false positives. Those whose health test results are negative are further defined as true negative.

The true positive ratio (TPR) and false positive ratio (FPR) obtained from each threshold were counted. For example, under the nth threshold, the true positive ratio is TPRn, and the false positive ratio is FPRn. The coordinate positions such as (FPR1, TPR1), (FPR2, TPR2), (FPR3, TPR3), etc. are sequentially drawn in the receiver operating characteristic graph. Among them, the abscissa of the receiver operating characteristic graph is a false positive ratio (FPR), and the ordinate is a true positive ratio (TPR). The inventor used the Analyse-it for Microsoft Excel software to plot the receiver operating characteristic curve.

The receiver operating characteristic curve is integrated to obtain the area under the curve (AUC) of the receiver, and the accuracy of the test can be evaluated. The closer the AUC value is to 1.0, the better the diagnostic accuracy is. The closer to 0, the worse the accuracy. If the AUC value is 1, the accuracy of the test is 100%. If the AUC value is 0, the accuracy of the detection is 0%. The detection method generally adopted by the market, the AUC is greater than 0.5.

B. Experimental results:

Please refer to Figure 1 and Table 1. Figure 1 is a receiver operating characteristic curve drawn from the SBI values of the schizophrenia group and its matching healthy group. As shown in Table 1, the area under the ROC curve is 0.8, and the 95% confidence interval is 0.71 to 0.89.

The above results show that the use of the linear acetylcholine receptor alpha alpha peptide linear polypeptide molecules in this case to detect schizophrenia, the area under the receiver operating characteristic curve is not only greater than 0.5, and up to 0.8. Therefore, the detection method of this case has considerable accuracy and application value.

Optimal threshold for linear peptide molecular assays for alpha 7-unit epitopes in neuroketamine acetylcholine receptors:

According to the distribution of each point on the receiver's operating characteristic curve, the optimal detection threshold is selected to make the detection method have high sensitivity (high sensitivity, ie, high true positive ratio), and high specificity (high specificity, ie, true negative ratio) .

Preferably, the optimal threshold of the detection method is SBI=2, that is, when the SBI value of the individual is higher than 2, the detection result is positive, and may be schizophrenia. Conversely, the individual's SBI is low. At 2 o'clock, the test result was negative, and the test did not find schizophrenia.

Please refer to Table 1. When SBI=2 was used as the threshold, the detection sensitivity of schizophrenia was 21.9% and the specificity was 90.7%.

It should be understood that the threshold corresponding to other points on the receiver operating characteristic curve may also be selected as a threshold for distinguishing between positive and negative.

The neuroic acetylcholine receptor alpha peptide unit linear polypeptide molecule of the invention can be used for preparing a diagnostic reagent or an immunozyme assay kit (ELISA Kit) for detecting autoantibody content in blood, serum or plasma of an individual. , Philip has diagnosed whether it is suffering from a mental illness. Preferably, it is used for detecting and diagnosing whether or not suffering from schizophrenia.

If the detected level of autoantibody is elevated, the function of the alpha 7 nicotine receptor representing the individual may be blocked/functioning, ie, the individual may have schizophrenia or neurocholine acetylcholine receptor 7 times. Other mental illnesses related to the unit.

An immunoenzyme assay kit prepared by using the alpha 7-element epitope linear polypeptide molecule of the neurogenic acetylcholine receptor of the present invention, comprising a porous disk, and the porous disk is coated with SEQ ID NO: 1, 2 , 3, 4, or 5 neuroacetylcholine receptor alpha alpha subunit epitope linear polypeptide molecule. Preferably, the porous disk is a 96-well disk. Preferably, the immunoenzyme assay kit further comprises reagents selected from the following tables:

The psychiatric diagnostic reagent or immunoenzyme assay kit prepared by using the linear 7-element epitope linear polypeptide molecule of the neurogenic acetylcholine receptor of the present invention can detect α 7 nAChR itself in blood, plasma, or serum of an individual. The antibody content is used as an objective basis for clinicians to comprehensively diagnose mental illness.

The above is only the preferred embodiment of the present invention, and is not intended to limit the scope of the claims of the present invention. Therefore, various other modifications or retouchings, etc., which are not departing from the spirit of the present invention, should be included in the present invention. Within the scope of the patent application.

The amino acid of the following sequence is represented from the left to the right in the direction of the amine group to the carboxyl group.

<110> Guanghui Life Medicine Co., Ltd.

<120> A polypeptide molecule for detecting an alpha subunit antibody of a neurocholine acetylcholine receptor

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<213> Artificial sequence

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<213> Artificial sequence

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Claims (10)

A mixture of antibody-binding molecules that binds to an antibody to one of the neuronal acetylcholine receptor subunit alpha-7 and can be used for early diagnosis of a mental illness or as a spirit of tracking An indicator of a disease condition; the antibody binding molecule mixture comprises: a first antibody binding molecule, the sequence of the first antibody binding molecule comprising SEQ ID NO: 1; a second antibody binding molecule, the sequence of the second antibody binding molecule comprising SEQ ID NO: 2; a third antibody binding molecule, the sequence of the third antibody binding molecule comprises SEQ ID NO: 3; a fourth antibody binding molecule, the sequence of the fourth antibody binding molecule comprises SEQ ID NO: 4; And a fifth antibody binding molecule comprising the sequence of SEQ ID NO: 5; wherein the first antibody binding molecule, the second antibody binding molecule, the third antibody binding molecule, the fourth antibody The binding characteristics of the binding molecule and the fifth antibody binding molecule further include: (a) the sequence length is 26-33 amino acids; (b) both are linear. Molecular; (c) the number of proline in the sequence is 0 to 3; (d) the number of glutamic acid in the sequence is 0 to 3; and (e) in the sequence The number of Cysteine is 0 to 1. The antibody binding molecule mixture according to claim 1, wherein the binding ability of the antibody binding molecule to the antibody is greater than the interaction between the antibody and a washing buffer. A detection reagent, which can be used for early diagnosis of a mental illness or as an indicator for tracking a mental illness condition; the detection reagent comprises: a first antibody binding molecule, the sequence of the first antibody binding molecule comprises SEQ ID NO: 1; a second antibody binding molecule, the sequence of the second antibody binding molecule comprising SEQ ID NO: 2; a third antibody binding molecule, the sequence of the third antibody binding molecule comprising SEQ ID NO: 3; a fourth antibody binding molecule The sequence of the fourth antibody binding molecule comprises SEQ ID NO: 4; and a fifth antibody binding molecule comprising the sequence of SEQ ID NO: 5; wherein the first antibody binding molecule, the first The sequence characteristics of the second antibody binding molecule, the third antibody binding molecule, the fourth antibody binding molecule, and the fifth antibody binding molecule further include: (a) the sequence length is 26-33 amino acids; (b ) are linear polypeptide molecules; (c) the number of proline in the sequence is 0 to 3; (d) the number of glutamic acid in the sequence is 0 to 3; e) Caspase in the sequence (Cysteine) are an acid number of 0-1. An immunoassay kit (ELISA Kit), which can be used for early diagnosis of a mental illness or as an indicator for tracking a mental illness condition. The immunoassay assay kit comprises a reagent, and the reagent further comprises: a first The antibody binding molecule, the sequence of the first antibody binding molecule comprises SEQ ID NO: 1; a second antibody binding molecule, the sequence of the second antibody binding molecule comprises SEQ ID NO: 2; a third antibody binding molecule, the The sequence of the triple antibody binding molecule comprises SEQ ID NO: 3; a fourth antibody binding molecule, the sequence of the fourth antibody binding molecule comprises SEQ ID NO: 4; and a fifth antibody binding molecule, the sequence of the fifth antibody binding molecule comprises SEQ ID NO: 5; Wherein, the sequence characteristics of the first antibody binding molecule, the second antibody binding molecule, the third antibody binding molecule, the fourth antibody binding molecule, and the fifth antibody binding molecule further comprise: (a) the length of the sequence 26-33 amino acids; (b) are linear polypeptide molecules; (c) the number of proline in the sequence is 0 to 3; (d) the glutamic acid in the sequence (Glutamic acid The number is 0 to 3; and the number of cysteine (Cysteine) in the (e) sequence is 0 to 1. The immunoenzyme assay kit according to claim 4, wherein the antibody binding molecule concentration in the reagent is 0.75 μg/mL to 5 mg/mL. An immunological enzyme assay kit (ELISA Kit) for early diagnosis of a mental illness or as an indicator for tracking a psychiatric condition; the immunoenzyme assay kit comprises a porous disk, and the porous disk comprises: a first antibody binding molecule, the sequence of the first antibody binding molecule comprising SEQ ID NO: 1; a second antibody binding molecule comprising a sequence comprising SEQ ID NO: 2; a third antibody binding molecule, The sequence of the third antibody binding molecule comprises SEQ ID NO: 3; a fourth antibody binding molecule comprising a sequence comprising SEQ ID NO: 4; and a fifth antibody binding molecule, the fifth antibody binding The sequence of the molecule comprises SEQ ID NO: 5; Wherein, the sequence characteristics of the first antibody binding molecule, the second antibody binding molecule, the third antibody binding molecule, the fourth antibody binding molecule, and the fifth antibody binding molecule further comprise: (a) the length of the sequence 26-33 amino acids; (b) are linear polypeptide molecules; (c) the number of proline in the sequence is 0 to 3; (d) the glutamic acid in the sequence (Glutamic acid The number is 0 to 3; and the number of cysteine (Cysteine) in the (e) sequence is 0 to 1. The immunoenzyme assay kit described in claim 4 or 6 is for detecting the antibody content of an alpha 7-unit unit of a neurotransmitter choline receptor in a blood, a plasma, or a serum. The antibody binding molecule mixture of claim 1, wherein the mental illness is schizophrenia. The test reagent of claim 3, wherein the mental illness is schizophrenia. The immunoenzyme assay kit of claim 4, wherein the mental illness is schizophrenia.