Tumor autoantigen / antibody combinations and uses for early detection of prostate cancer

Abstract

The present application relates to tumor autoantigen / antibody combinations and applications for early detection of prostate cancer. Specifically, the present application provides the use of diagnostic reagents for autoantibodies against target antigens. The research of the present application demonstrates that the autoantibody combinations of the present application can be used for early screening or differential diagnosis of prostate cancer, as well as for efficacy evaluation after intervention treatment of prostate cancer patients. The detection method of the present application is simple and fast, has high patient compliance, high sensitivity and high specificity, thereby helping to more accurately and earlier diagnose and prognostic evaluate prostate cancer.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biotechnology and medical diagnostics, and in particular, the present application relates to tumor autoantigen / antibody combinations and uses for early detection of prostate cancer. BACKGROUND

[0002] According to the International Agency for Research on Cancer of the World Health Organization, 1.1 million men are diagnosed with prostate cancer worldwide each year, accounting for 15% of all cancers diagnosed in men. An estimated 307,000 people die from prostate cancer worldwide each year, making it the fifth leading cause of cancer death in men. The estimated new cases of prostate cancer in China are about 72,000, and the incidence rate is 6.59 / 100,000 in the Chinese standard population, which is similar to the world standard population, ranking sixth in the male malignant tumor incidence spectrum; the estimated death cases are about 30,700, and the mortality rate is 2.61 / 100,000 in the Chinese standard population, ranking tenth in the male malignant tumor mortality spectrum.

[0003] Although the exact cause of prostate cancer is not known, the main risk factor for prostate cancer is advanced age. About 75% of prostate cancers are diagnosed in men over 65 years old. Prostate cancer is rarely seen before the age of 50. In addition to advanced age, race is considered a risk factor for prostate cancer, although the exact association is not very clear. There is a close association between family history and the risk of prostate cancer. Men with a first-degree relative (e.g., father, brother, son) with prostate cancer are two to three times more likely to be diagnosed with prostate cancer, and this risk increases with the number of relatives diagnosed with prostate cancer.

[0004] Prostate-specific antigen (PSA) is the most common test for prostate cancer. Given that there are many factors that can cause PSA to rise, it is often difficult to explain the clear reason for the rise in PSA. Prostate cancer PSA screening has also raised concerns about the risk of unnecessary biopsy, overdiagnosis and overtreatment of indolent tumors detected by screening, and possible adverse side effects. Therefore, PSA is difficult to be a universally applicable, stable, and highly specific marker for prostate cancer.

[0005] The Prostate Health Index (PHI) is a blood test that estimates the risk of prostate cancer. The PHI test is a combination of free PSA, total PSA, and the [-2] proPSA isoform of free PSA. The three test molecules are in a formula that calculates a PHI score. The PHI score is a better predictor of prostate cancer than a total PSA test alone or a free PSA test alone. Nevertheless, PHI cannot escape the inherent shortcomings of PSA as a marker for prostate cancer.

[0006] Another type of early detection test for prostate cancer is a digital rectal exam (DRE). DRE focuses on the size and consistency of the prostate. Many cancers occur in the peripheral zone of the prostate, so they can be detected by a DRE. Cancers are characterized as hard, nodular, and irregular. During a DRE, the clinician’s finger palpates the posterior and lateral aspects of the prostate. Limitations of DRE include the inability to palpate tumors in the anterior and midline regions. Overall, approximately 25% of men with an abnormal DRE have cancer, so the specificity of DRE is also not ideal. In addition, DRE causes pain or discomfort in most patients, making it difficult to use widely.

[0007] Prostate cancer antigen 3 (PCA3) is a non-coding RNA that is overexpressed in prostate cancer tissue. This marker is measured by analyzing expression in prostate cells from a urine sample after a vigorous prostate massage. Currently, there is insufficient data to support the use of urine PCA3 testing for prostate cancer screening, but it can help determine whether a repeat biopsy is needed.

[0008] Multiparametric magnetic resonance imaging of the prostate has been shown to be a means of detecting and further characterizing prostate tumors. This technique can be used for men who require monitoring of a persistent rise in PSA, as well as for patients with a rising PSA and a benign lesion on biopsy. While multiparametric magnetic resonance imaging can be used to identify targets for monitoring prostate lesions, it cannot replace simple procedures that are easy to promote for screening or pathological biopsy for diagnosis, and there is a lack of large-scale randomized trials, as well as high cost of detection.

[0009] The ultimate goal of screening for prostate cancer should be to reduce the incidence and suffering of advanced disease, and not to harm men who do not need treatment for low-risk disease. Diagnosing cancer based on serum analysis is a particularly attractive concept. One potential biomarker strategy for identifying cancer is to take advantage of the body’s own immune system. Cancer sera contain a unique set of antibodies that react with self-cell antigens, known as tumor-associated autoantibodies (TAAB). Proteins that are not present in normal cells and are abnormal can trigger a host immune response, and the form of TAAB antibodies is often significantly amplified relative to the small amount of antigen. Since tumor immune responses are locally produced, even a small amount of antigen can not be detected by any other means, and can be recognized and amplified by the immune response, especially in the early stages of cancer formation. Therefore, autoantibody analysis can be an effective method for identifying cancer. The current antibody molecule combination strategy for detecting prostate cancer still has the problem of insufficient specificity, such as the combination of TARDBP, TLN1, PARK7, LEDGF / PSIP1, and CALD1, which has a detection specificity of 80% for healthy people. Such problems can mislead patients about prostate cancer and make it difficult to translate to the clinic.

[0010] Therefore, there is a lack of a molecular combination of autoantibodies for prostate cancer in the current art to assist in the diagnosis of the disease, and it is still necessary to find new combinations of autoantibody biomarkers.

[0011] Therefore, it is the object of the present application to provide a simple, efficient, highly sensitive and specific combination of biomarkers for the detection of prostate cancer. SUMMARY

[0012] The present application provides the use of a combination of tumor autoantigens / antibodies for the early detection of prostate cancer in serum / plasma samples.

[0013] In a first aspect of the present application, there is provided the use of a diagnostic agent for an autoantibody to a target antigen for the manufacture of a diagnostic agent or kit for (a) diagnosing the risk of developing prostate cancer; and / or (b) prognostic evaluation of prostate cancer;

[0014] wherein the autoantibody to a target antigen is selected from the group consisting of:

[0015] (A) any antibody selected from the group consisting of A1 to A8, or a combination thereof: (A1) Anti-COPB1; (A2) Anti-EIF3E; (A3) Anti-CCDC110; (A4) Anti-MAGEA4; (A5) Anti-HNRPA1; (A6) Anti-HMGB3; (A7) Anti-C1D; (A8) Anti-BRCA2.

[0016] In another preferred embodiment, the autoantibody to a target antigen further comprises (B) any marker selected from the group consisting of B1 to B2, or a combination thereof: (B1) Anti-P53; (B2) Anti-BRCA1.

[0017] In another preferred embodiment, the autoantibody to a target antigen comprises at least 2 markers selected from the group consisting of A1 to A8.

[0018] In another preferred embodiment, the autoantibody to a target antigen comprises 3, 4, 5, 6, 7 or 8 markers selected from the group consisting of A1 to A8.

[0019] In another preferred embodiment, the autoantibody to a target antigen comprises a combination of one or more markers selected from the group consisting of A1 to A8 and one or more markers selected from the group consisting of B1 to B2.

[0020] In another preferred embodiment, the combination of autoantibodies against target antigens is: (Al) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (B 1) Anti-P53, and (B2) Anti-BRCA1.

[0021] In another preferred embodiment, the combination of autoantibodies against target antigens is: (Al) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (B 1) Anti-P53, and (B2) Anti-BRCA1.

[0022] In another preferred embodiment, the combination of autoantibodies against target antigens is: (Al) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (B 1) Anti-P53, and (B2) Anti-BRCA1.

[0023] In another preferred embodiment, the combination of autoantibodies against target antigens is: (Al) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (B 1) Anti-P53, and (B2) Anti-BRCA1.

[0024] In another preferred embodiment, the combination of autoantibodies against target antigens is: (Al) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (B 1) Anti-P53, and (B2) Anti-BRCA1.

[0025] In another preferred embodiment, the combination of autoantibodies against target antigens is: (Al) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (B 1) Anti-P53, and (B2) Anti-BRCA1.

[0026] In another preferred embodiment, the combination of autoantibodies against target antigens is: (Al) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (B 1) Anti-P53, and (B2) Anti-BRCA1.

[0027] In another preferred embodiment, the combination of autoantibodies against target antigens is: (Al) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (B 1) Anti-P53, and (B2) Anti-BRCA1.

[0028] In another preferred embodiment, the combination of autoantibodies against target antigens is: (Al) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (B 1) Anti-P53, and (B2) Anti-BRCA1.

[0029] In another preferred embodiment, the autoantibodies against target antigens are combined as follows: (A1) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (A6) Anti-HMGB3, (Bl) Anti-P53, and (B2) Anti-BRCA1.

[0030] In another preferred embodiment, the autoantibodies against target antigens are combined as follows: (A1) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (A6) Anti-HMGB3, (Bl) Anti-P53, and (B2) Anti-BRCA1.

[0031] In another preferred embodiment, the autoantibodies against target antigens are combined as follows: (A1) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (A6) Anti-HMGB3, (Bl) Anti-P53, and (B2) Anti-BRCA1.

[0032] In another preferred embodiment, the autoantibodies against target antigens are combined as follows: (A1) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (A6) Anti-HMGB3, (Bl) Anti-P53, and (B2) Anti-BRCA1.

[0033] In another preferred embodiment, the autoantibodies against target antigens are combined as follows: (A1) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (A6) Anti-HMGB3, (Bl) Anti-P53, and (B2) Anti-BRCA1.

[0034] In another preferred embodiment, the autoantibodies against target antigens are combined as follows: (A1) Anti-COPBl, (A2) Anti-EIF3E, (A3) Anti-CCDC110, (A4) Anti-MAGEA4, (A6) Anti-HMGB3, (Bl) Anti-P53, and (B2) Anti-BRCA1.

[0035] In another preferred embodiment, the autoantibodies against the target antigens are combined as: (Al) Anti-COPBl, (A2) Anti-EIF3E, (Bl) Anti-P53, (A3) Anti-CCDC110, (B2) Anti-BRCA1, (A4) Anti-MAGEA4, (A6) Anti-HMGB3, (A7) Anti-C1D, (A8) Anti-BRCA2, and (A5) Anti-HNRPA1.

[0036] In another preferred embodiment, the diagnostic reagent of the autoantibodies against the target antigens is an antigen protein (as a positive control).

[0037] In another preferred embodiment, the antigen protein is selected from the group consisting of:

[0038] (C) any antigen selected from the group C1-C10, or a combination thereof: (C1) COPBl; (C2) EIF3E; (C3) CCDC110; (C4) MAGEA4; (C5) HNRPA1; (C6) HMGB3; (C7) C1D; (C8) BRCA2; (C9) P53; (C10) BRCA1.

[0039] In another preferred embodiment, the antigen protein is: (C1) COPBl, (C2) EIF3E, (C9) P53, and (C3) CCDC110.

[0040] In another preferred embodiment, the antigen protein is: (C10) BRCA1, (C4) MAGEA4, and (C5) HNRPA1.

[0041] In another preferred embodiment, the antigen protein is: (C6) HMGB3, (C7) C1D, and (C8) BRCA2.

[0042] In another preferred embodiment, the antigen protein is: (C1) COPBl and (C2) EIF3E.

[0043] In another preferred embodiment, the antigen protein is: (C1) COPBl, (C2) EIF3E, and (C9) P53.

[0044] In another preferred embodiment, the antigen protein is: (C1) COPBl, (C2) EIF3E, (C9) P53, (C3) CCDC110, and (C10) BRCA1.

[0045] In another preferred embodiment, the antigen protein is: (C1) COPBl, (C2) EIF3E, (C9) P53, (C3) CCDC110, and (C4) MAGEA4.

[0046] In another preferred example, the antigen proteins are: (C1) COPBl, (C2) EIF3E, (C9) P53, (C3) CCDC110, (C10) BRCA1, (C4) MAGEA4, and (C6) HMGB3.

[0047] In another preferred example, the antigen proteins are: (C10) BRCA1, (C4) MAGEA4, (C6) HMGB3, (C7) C1D, (C8) BRCA2, and (C5) HNRPA1.

[0048] In another preferred example, the antigen proteins are: (C1) COPBl, (C2) EIF3E, (C9) P53, (C3) CCDC110, (C10) BRCA1, (C4) MAGEA4, and (C6) HMGB3.

[0049] In another preferred example, the antigen proteins are: (C1) COPBl, (C2) EIF3E, (C9) P53, (C3) CCDC110, (C10) BRCA1, (C4) MAGEA4, and (C7) C1D.

[0050] In another preferred example, the antigen proteins are: (C1) COPBl, (C2) EIF3E, (C9) P53, (C3) CCDC110, (C10) BRCA1, (C4) MAGEA4, and (C8) BRCA2.

[0051] In another preferred example, the antigen proteins are: (C1) COPBl, (C2) EIF3E, (C9) P53, (C3) CCDC110, (C10) BRCA1, (C4) MAGEA4, and (C5) HNRPA1.

[0052] In another preferred example, the antigen proteins are: (C1) COPBl, (C2) EIF3E, (C9) P53, (C3) CCDC110, (C6) HMGB3, (C7) C1D, (C8) BRCA2, and (C5) HNRPA1.

[0053] In another preferred example, the antigen proteins are: (C1) COPBl, (C2) EIF3E, (C9) P53, (C3) CCDC110, (C6) HMGB3, (C7) C1D, (C8) BRCA2, and (C5) HNRPA1.

[0054] In another preferred embodiment, the antigen proteins are: (C1) COPB1, (C2) EIF3E, (C9) P53, (C3) CCDC110, (C10) BRCA1, (C4) MAGEA4, (C6) HMGB3, (C7) C1D, (C8) BRCA2 and (C5) HNRPA1.

[0055] In a second aspect of the present application, a kit is provided, which comprises a detection reagent for detecting autoantibodies against target antigens,

[0056] In another preferred embodiment, the autoantibodies against target antigens are selected from the group consisting of:

[0057] (A) a combination of two or more autoantibodies selected from A1 to A8:

[0058] (A1) Anti-COPB1; (A2) Anti-EIF3E; (A3) Anti-CCDC110; (A4) Anti-MAGEA4; (A5) Anti-HNRPA1; (A6) Anti-HMGB3; (A7) Anti-C1D; (A8) Anti-BRCA2.

[0059] In another preferred embodiment, the autoantibodies against target antigens further comprise:

[0060] (B) a combination of one or more autoantibodies selected from A1 to A8 and one or more autoantibodies selected from B1 to B2:

[0061] In another preferred embodiment, the autoantibodies against target antigens further comprise:

[0062] In another preferred embodiment, the kit detects the autoantibodies against target antigens through antigen-antibody reaction.

[0063] In another preferred embodiment, the kit detects the autoantibodies against target antigens through antigen-antibody reaction.

[0064] In a third aspect of the present application, a detection method is provided, comprising the steps of:

[0065] (a) providing a detection sample;

[0066] (b) detecting the level of autoantibodies against target antigens in the detection sample, denoted as Y1; and

[0067] (c) comparing the level of autoantibodies against target antigens with a control reference value Y0.

[0068] wherein the autoantibody against the target antigen is selected from the group consisting of:

[0069] (A) any antibody selected from the group consisting of A1 to A8, or a combination thereof: (A1) Anti-COPB1; (A2) Anti-EIF3E; (A3) Anti-CCDC110; (A4) Anti-MAGEA4; (A5) Anti-HNRPA1; (A6) Anti-HMGB3; (A7) Anti-C1D; (A8) Anti-BRCA2;

[0070] If the detection result of the autoantibody against the target antigen of the detection subject meets the following condition, it is suggested that the detection subject has a high risk of prostate cancer:

[0071] When the level of a certain autoantibody is higher than the reference value or standard value Y0, it is suggested that the detection subject has a high risk of prostate cancer.

[0072] In another preferred embodiment, the autoantibody against the target antigen further comprises:

[0073] (B) a combination of one or more autoantibodies selected from the group consisting of A1 to A8 and one or more autoantibodies selected from the group consisting of B1 to B2.

[0074] wherein the autoantibodies B1 to B2 are: (B1) Anti-P53; (B2) Anti-BRCA1.

[0075] In another preferred embodiment, the detection sample is selected from the group consisting of whole blood, serum, plasma, tissue, cells, intercellular fluid, cerebrospinal fluid, urine, or a combination thereof.

[0076] In another preferred embodiment, the detection sample is selected from the group consisting of whole blood, serum, and plasma.

[0077] In a fourth aspect of the present application, a prostate cancer risk determination device is provided, the device comprising:

[0078] (a) an input module for inputting autoantibody data against a target antigen of a subject;

[0079] wherein the autoantibody against the target antigen is selected from the group consisting of:

[0080] (A) any one antibody selected from the group consisting of A1 to A8, or a combination thereof: (A1) Anti-COPBl; (A2) Anti-EIF3E; (A3) Anti-CCDC110; (A4) Anti-MAGEA4; (A5) Anti-HNRPA1; (A6) Anti-HMGB3; (A7) Anti-ClD; (A8) Anti-BRCA2;

[0081] (b) a processing module, which compares the inputted autoantibody level Y1 with the control reference value Y0, thereby obtaining a judgment result, wherein when the comparison result meets a judgment condition, it is suggested that the subject has a high risk of prostate cancer; otherwise, it is suggested that the subject does not have a high risk of prostate cancer;

[0082] (c) an output module, which outputs the judgment result.

[0083] In another preferred embodiment, the autoantibody against the target antigen further comprises:

[0084] (B) a combination of one or more autoantibodies selected from A1 to A8 and one or more autoantibodies selected from B1 to B2.

[0085] The autoantibodies B1 to B2 are: (B1) Anti-P53; (B2) Anti-BRCA1.

[0086] In a fifth aspect of the present application, a use of an autoantibody-antigen combination diagnostic reagent for preparing a diagnostic reagent or kit for jointly diagnosing the occurrence risk of prostate cancer is provided.

[0087] The autoantibody-antigen combination comprises: an autoantigen PSA; and

[0088] an autoantibody selected from the group consisting of:

[0089] (A) any one antibody selected from the group consisting of A1 to A8, or a combination thereof: (A1) Anti-COPBl; (A2) Anti-EIF3E; (A3) Anti-CCDC110; (A4) Anti-MAGEA4; (A5) Anti-HNRPA1; (A6) Anti-HMGB3; (A7) Anti-ClD; (A8) Anti-BRCA2.

[0090] In another preferred embodiment, the autoantibody further comprises:

[0091] (B) a combination of one or more autoantibodies selected from A1 to A8 and one or more autoantibodies selected from B1 to B2.

[0092] The B1 to B2 autoantibodies are: (B1) Anti-P53; (B2) Anti-BRCA1.

[0093] In another preferred embodiment, the diagnostic reagent for the autoantigen PSA is: Anti-PSA.

[0094] In another preferred embodiment, the antibody-antigen combination is: the autoantigen PSA and the following seven autoantibodies: (A1) Anti-COPB1; (A2) Anti-EIF3E; (A3) Anti-CCDC110; (A4) Anti-MAGEA4; (A5) Anti-HNRPA1; (B1) Anti-P53; (B2) Anti-BRCA1.

[0095] It should be understood that, within the scope of the present application, the above technical features of the present application and the technical features specifically described below (e.g., in the examples) can be combined with each other to form new or preferred technical solutions. Due to the limited space, they will not be listed one by one here. BRIEF DESCRIPTION OF DRAWINGS

[0096] Figure 1 The scatter plots of the levels of each autoantibody in the prostate cancer group, the benign disease group, and the healthy control group are shown.

[0097] Figure 2 The ROC curve of the preferred autoantibody combination of the present application (Anti-COPB1, Anti-EIF3E, Anti-P53, Anti-CCDC110, Anti-BRCA1, Anti-MAGEA4, Anti-HNRPA1) in distinguishing the healthy controls and the prostate cancer patients in this study is shown.

[0098] Figure 3 The ability of the preferred autoantibody combination of the present application (Anti-COPB1, Anti-EIF3E, Anti-P53, Anti-CCDC110, Anti-BRCA1, Anti-MAGEA4, Anti-HNRPA1) in distinguishing the benign disease and the prostate cancer patients in this study is shown.

[0099] Figure 4 The detection ability of the detection model of the present application (Anti-COPB1, Anti-EIF3E, Anti-P53, Anti-CCDC110, Anti-BRCA1, Anti-MAGEA4, Anti-HNRPA1) for prostate cancer of different pathological grades (Gleason score) is shown.

[0100] Figure 5The detection ability of the inventive autoantibody panel (Anti-COPBl, Anti-EIF3E, Anti-P53, Anti-CCDC110, Anti-BRCA1, Anti-MAGEA4, Anti-HNRPA1) for different pathological grades of prostate cancer is shown.

[0101] Figure 6 The ROC plot of the inventive autoantibody panel (Anti-COPBl, Anti-EIF3E, Anti-P53, Anti-CCDC110, Anti-BRCA1, Anti-MAGEA4, Anti-HNRPA1) for low PSA prostate cancer is shown.

[0102] Figure 7 The ROC plot of the inventive autoantibody panel (Anti-COPBl, Anti-EIF3E, Anti-P53, Anti-CCDC110, Anti-BRCA1, Anti-MAGEA4, Anti-HNRPA1) for high PSA prostate cancer is shown at the maximum of the Youden index.

[0103] Figure 8 The ROC plot of PSA alone as a marker for prostate cancer in discriminating healthy individuals from prostate cancer patients is shown.

[0104] Figure 9 The ROC plot of PSA alone as a marker for prostate cancer in discriminating benign disease patients from prostate cancer patients is shown.

[0105] Figure 10 The ROC plot of the inventive preferred autoantibody panel in combination with PSA (i.e. the molecular combination Anti-COPBl, Anti-EIF3E, Anti-P53, Anti-CCDC110, Anti-BRCA1, Anti-MAGEA4, Anti-HNRPA1 and PSA) for the ability to discriminate between healthy controls and prostate cancer patients in this study is shown.

[0106] Figure 11 The ROC plot of the inventive preferred autoantibody panel in combination with PSA (i.e. the molecular combination Anti-COPBl, Anti-EIF3E, Anti-P53, Anti-CCDC110, Anti-BRCA1, Anti-MAGEA4, Anti-HNRPA1 and PSA) for the ability to discriminate between benign disease and prostate cancer patients in this study is shown.

[0107] Figure 12 The comparison of the sensitivity of different autoantibody panels of the invention is shown.

[0108] Figure 13 The specificity of different combinations of the antibodies of the present application is shown. DETAILED DESCRIPTION

[0109] The present inventors have made extensive and in-depth studies and unexpectedly found a high-sensitivity and high-specificity autoantibody against target antigens for the diagnosis and prognosis of prostate cancer. The autoantibody against target antigens for the diagnosis and prognosis of prostate cancer comprises a total of 10 autoantibodies selected from Group A and Group B, and a method and kit for evaluating the risk and prognosis of prostate cancer are accordingly developed. The autoantibody can also be combined with the autoantigen PSA to judge the risk and prognosis of prostate cancer, and can detect low-PSA (≤4 ng / mL) prostate cancer patients with high sensitivity and specificity. On this basis, the present application is completed.

[0110] TERMS

[0111] In the present application, the term "antigen" or the term "antigen protein" can be used interchangeably.

[0112] The terms "antibody", "autoantibody", "autoantibody of the present application", and "autoantibody of the present application against target antigens" can be used interchangeably in the present application.

[0113] In addition, the following experimental operations or definitions are involved in the present application. It should be noted that the present application can also be implemented using other conventional techniques in the art, and is not limited only to the following experimental operations.

[0114] (I) Preparation of recombinant antigen protein

[0115] The cDNA fragment of the tumor antigen is cloned into a PET28(a) expression vector containing a 6XHis tag. A streptavidin protein or the like (a tag protein that binds to biotin) is introduced at the N- or C-terminus of the antigen. The obtained recombinant expression vector is transformed into E. coli for expression. The expressed protein in the supernatant is purified by a Ni-NTA affinity column and an ion column. When the protein is expressed in an inclusion body, the protein is denatured with 6M guanidine hydrochloride and refolded in vitro according to the standard method, and then purified by a Ni-NTA affinity column through the 6XHis tag to obtain the antigen protein.

[0116] (II) Preparation and preservation of serum or plasma

[0117] The serum or plasma of a prostate cancer patient is collected when the patient is initially diagnosed with prostate cancer and has not yet received any radiotherapy, chemotherapy, and surgical treatment. The plasma or serum is prepared according to the standard clinical procedure and stored in a -80°C refrigerator for long-term preservation.

[0118] (III) ELISA detection

[0119] The concentration of autoantibody markers in the sample is quantified by enzyme-linked immunosorbent assay (ELISA). Purified tumor antigens are immobilized on the surface of micro-wells via their tag, streptavidin or analog. The micro-wells are pre-coated with biotin-labeled bovine serum albumin (BSA). Serum or plasma samples are diluted 1:110 in phosphate buffer and added to the micro-wells for reaction (50 μl / well). After washing to remove unbound serum or plasma components, horseradish peroxidase (HRP)-conjugated anti-human IgG is added to each well for reaction. The reaction substrate TMB (3,3',5,5'-tetramethylbenzidine) is then added for color development. A stop solution (1 N HCl) is added, and the absorbance is read at 450 nm single wavelength on a microplate reader (OD). The serum autoantibody concentration is quantified using a standard curve.

[0120] The concentration of antigen markers in the sample is quantified by sandwich enzyme-linked immunosorbent assay. Specific antibodies are linked to a solid phase carrier to form a solid phase antibody, and unbound antibodies and impurities are removed by washing. The sample to be tested, i.e., serum or plasma, is diluted 1:110 in phosphate buffer and added to the micro-wells for reaction (50 μl / well), allowing it to react with the solid phase antibody for a period of time to allow the antigen in the sample to bind to the antibody on the solid phase carrier to form a solid phase antigen complex. Other unbound substances are removed by washing. Horseradish peroxidase (HRP)-conjugated anti-human IgG is added for reaction. The reaction substrate TMB (3,3',5,5'-tetramethylbenzidine) is then added for color development. A stop solution (1 N HCl) is added, and the absorbance is read at 450 nm single wavelength on a microplate reader (OD). At this time, the amount of enzyme carried on the solid phase carrier is directly related to the amount of the substance to be tested in the sample. The enzyme in the sandwich complex catalyzes the substrate to become a colored product. Qualitative or quantitative determination of the antigen is performed according to the degree of color reaction.

[0121] (iv) Cutoff value of autoantibody and / or antigen protein

[0122] The cutoff value of autoantibody and / or antigen level is defined as equal to the average value of the healthy control cohort in the control group (the control group is a population confirmed by physical examination to be free of cancer) plus 2 standard deviations (SD).

[0123] (v) Positive and negative judgment of individual autoantibody and antigen protein

[0124] For each autoantibody and / or antigen protein determination, a positive reaction is defined as the level of autoantibody and / or antigen protein in the sample is quantified and compared with the cutoff value, and ≥ the cutoff value is positive; correspondingly, a negative reaction is defined as < the cutoff value is negative.

[0125] (VI) Positive judgment of the combination of autoantibodies and / or antigen proteins

[0126] Since the positive rate of a single autoantibody and / or a single antigen protein is low, in order to increase the positive rate of the detection of autoantibodies and / or antigen proteins, the results of multiple autoantibodies and / or multiple antigen proteins are combined when analyzing the results to judge the prediction effect.

[0127] The rules are:

[0128] (1) Multiple autoantibodies are detected in the sample, and as long as one or more of the autoantibodies shows positive, the antibody combination result is judged to be positive; and if all the autoantibodies are negative, the result is judged to be negative.

[0129] (2) Multiple antigen proteins are detected in the sample, and as long as one or more of the antigen proteins shows positive, the result is judged to be positive; and if all the antigen proteins are negative, the result is judged to be negative.

[0130] (3) Multiple autoantibodies and multiple antigen proteins are detected in the sample at the same time, and as long as one or more of the autoantibodies and / or antigen proteins is positive, the result is judged to be positive; and if all the antibodies and antigen proteins are negative, the result is judged to be negative.

[0131] (VII) Statistical analysis method

[0132] Statistical analysis was performed on two groups using Mann-Whitney U test using GraphPad Prism v.6 (Graphpad Prism software, San Diego, CA) and IBM SPSS Statistics 23 for Windows (IBM, New York, New York). Spearman's correlation analysis was performed when analyzing the relationship between each parameter.

[0133] (VIII) Sensitivity and specificity judgment

[0134] Sensitivity: The proportion of cases in which the results of autoantibodies, autoantibody combinations, antigen proteins, antigen protein combinations, and autoantibody and antigen protein combinations are positive among all cases diagnosed as prostate cancer by the gold standard.

[0135] Specificity: The proportion of subjects in which the results of autoantibodies, autoantibody combinations, antigen proteins, antigen protein combinations, and autoantibody and antigen protein combinations are negative among all subjects diagnosed as disease-free by the gold standard.

[0136] Prostate-specific antigen (PSA)

[0137] Prostate specific antigen (PSA) is the most common test for prostate cancer. PSA is a serine protease, a member of the kallikrein family of enzymes encoded on chromosome 13, and is present in the luminal epithelial cells of the prostate ducts. Anything that disrupts the natural tissue layers of the prostatic ducts can cause an increase in PSA. Non-malignant causes of elevated PSA in serum include benign prostatic hyperplasia (BPH), prostatitis, urinary retention, or instrumentation (e.g., urinary catheterization, transrectal ultrasound, and prostate biopsy). Given the number of factors that can cause an increase in PSA, it is often difficult to explain the clear cause of an elevated PSA. Furthermore, another cause of the confusing interpretation of serum PSA levels is the controversial "normal" value. PSA is measured in nanograms per milliliter (ng / mL), and the accepted normal reference range is 0 to 4 ng / mL, at which time the specificity of using PSA as a marker for prostate cancer is only 60%.

[0138] Because PSA levels rise with age and prostate volume (i.e., BPH), it is recommended that age-adjusted values be used. In addition, racial differences also cause variations in the PSA "normal" range. Prostate cancer PSA screening has also raised concerns about the risk of unnecessary biopsies, overdiagnosis of indolent tumors and overtreatment, and possible adverse side effects. Thus, PSA is difficult to be a universally applicable, stable, and highly specific marker for prostate cancer.

[0139] Detection methods

[0140] Based on the elevated levels of autoantibodies against target antigens in blood, plasma or serum, etc. in prostate patients, the present application also provides corresponding methods for diagnosing the risk of prostate disease.

[0141] The autoantibodies against target antigens provided by the present application include a total of 10 autoantibodies selected from Group A and selected from Group B: (A1) Anti-COPB1; (A2) Anti-EIF3E; (A3) Anti-CCDC110; (A4) Anti-MAGEA4; (A5) Anti-HNRPA1; (A6) Anti-HMGB3; (A7) Anti-C1D; (A8) Anti-BRCA2; (B1) Anti-P53; (B2) Anti-BRCA1.

[0142] The present application relates to diagnostic test methods for quantitatively and qualitatively detecting the levels of autoantibodies against target antigens in humans. These tests are well known in the art. The levels of autoantibodies against target antigens detected in the tests can be used to diagnose (including auxiliary diagnosis) the risk of the occurrence of prostate cancer, and / or the prognosis evaluation of prostate cancer.

[0143] A preferred method is to quantitatively detect autoantibodies against the target antigen.

[0144] Preferably, a method for detecting whether there is an autoantibody against the target antigen in a sample is to use specific antigens for detection, which comprises: contacting the sample with an antigen protein specific antibody; observing whether an antibody complex is formed, and if the antibody complex is formed, it indicates that there is an autoantibody against the target antigen in the sample.

[0145] The autoantibodies against the target antigen of the present application can be used for the diagnosis of prostate cancer. The antigen protein of the autoantibodies against the target antigen can be fixed on a protein chip for detecting the autoantibodies against the target antigen in a sample.

[0146] Based on the research of the present application, the level of the autoantibodies against the target antigen of the present application is significantly increased in prostate cancer patients. Therefore, the autoantibodies against the target antigen of the present application can be used as a marker for detecting or diagnosing (especially auxiliary diagnosis and / or early diagnosis) the risk of prostate cancer. In detection, if the ratio of the level of the autoantibodies Y1 to the corresponding level Y0 in the normal population (Y1 / Y0) is ≥1.5, preferably ≥2, more preferably ≥3, it can be considered as an increased risk of prostate cancer.

[0147] In addition, the present inventors have unexpectedly found that the preferred combination of autoantibodies against the target antigen of the present application has better detection ability for prostate cancer patients with higher pathological grade (Gleason score), and has better detection rate for prostate cancer patients with medium and low PSA levels.

[0148] Detection kit

[0149] Based on the correlation between the autoantibodies against the target antigen of the present application and the risk and prognosis of prostate cancer, the autoantibodies against the target antigen of the present application can be used as a diagnostic marker for the occurrence of prostate cancer, and / or a prognostic evaluation marker for prostate cancer.

[0150] The autoantibodies against the target antigen provided by the present application include a total of 10 autoantibodies selected from group A and selected from group B: (A1) Anti-COPB1; (A2) Anti-EIF3E; (A3) Anti-CCDC110; (A4) Anti-MAGEA4; (A5) Anti-HNRPA1; (A6) Anti-HMGB3; (A7) Anti-C1D; (A8) Anti-BRCA2; (B1) Anti-P53; (B2) Anti-BRCA1.

[0151] The present application also provides a kit for diagnosing the occurrence of prostate cancer, which comprises a detection reagent for detecting the autoantibody against the target antigen according to the present application. Preferably, the kit comprises the antigen of the autoantibody against the target antigen according to the present application, or an active fragment thereof.

[0152] In another preferred embodiment, the kit further comprises a label or an instruction indicating that the kit is used for diagnosing the risk of occurrence of prostate cancer and / or evaluating the prognosis of prostate cancer.

[0153] The main advantages of the present application are:

[0154] (1) The diagnosis or detection of prostate cancer by the autoantibody according to the present application only needs serum or plasma sample, without the need of taking tissue sample for detection, which avoids the resistance of patients to such detection, helps to find early prostate cancer patients, prolongs the survival period of patients and improves the quality of life, and has a high clinical application prospect.

[0155] (2) The autoantibody combination according to the present application has a good distinguishing ability for prostate cancer patients and prostate benign diseases.

[0156] (3) Compared with the traditional tumor markers or the detection or diagnosis of prostate cancer by a single autoantibody according to the present application, the sensitivity of the autoantibody according to the present application or the combination of the autoantibody and PSA is significantly improved.

[0157] (4) The autoantibody combination according to the present application has excellent detection ability for prostate cancer patients with low and medium PSA levels which cannot be effectively detected in the prior art, and can further reduce the harm caused by prostate cancer.

[0158] (5) The autoantibody combination according to the present application has better detection ability for prostate cancer with high malignancy than for prostate cancer with low malignancy, which helps to diagnose and treat as early as possible and prolong the survival period of patients.

[0159] The present application will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present application and not used to limit the scope of the present application. The experimental methods in the following examples without specific conditions are usually carried out according to the conventional conditions, such as the conditions described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are weight percentages and weight parts.

[0160] Example 1

[0161] Example 1. Single autoantibody sensitivity and specificity detection in patient serum samples

[0162] This example includes 29 healthy subjects, 46 benign prostate disease patients and 105 prostate cancer patients for autoantibody marker screening. The subjects are from at least 3 different medical centers. All the prostate cancer patient sera are collected at the time of diagnosis before any radiotherapy, chemotherapy or surgery treatment, and stored at -80°C. The subject information is shown in Table 1.

[0163] Table 1

[0164]

[0165] Prostate cancer antigens are coated on the surface of 96-well plates after expression and purification, and then reacted with 1:110 diluted prostate cancer serum, benign disease patient serum or healthy control serum after blocking. Then, the plates are reacted with anti-human IgG antibody-HRP, followed by color development, and detected by an enzyme-labeled instrument at 450 nm wavelength. The detection sensitivity and specificity are shown in Table 2.

[0166] Table 2. Sensitivity and specificity of single autoantibody as a marker in serum samples

[0167]

[0168]

[0169] The scatter plot of the level distribution of the above 10 autoantibodies in the prostate cancer group, benign disease group and healthy control group is shown in Figure 1 and Table 3. Due to the differences in the immune system of tumor patients and the diversity of tumor generation mechanisms, the sensitivity of single tumor autoantibody in tumor patients is low.

[0170] Table 3. Level distribution data table of 10 autoantibodies in prostate cancer group, benign disease group and healthy control group (unit: U / mL)

[0171]

[0172] Note: U / mL (unit / ml), antibody titer is used to measure the minimum concentration (i.e. maximum dilution) required for an antibody to recognize a specific epitope. It is generally expressed as: the maximum dilution that can still produce a positive result. Antibody titer is usually detected by ELISA method.

[0173] Statistical analysis of the level distribution of autoantibodies in the prostate cancer group, benign disease group and healthy control group was performed using the Kruskal-Wallis test, and it was found that the antibodies against COPB1, EIF3E, P53, HMGB3, C1D, BRCA2 and HNRPA1 in the prostate cancer group were significantly higher than those in the benign disease group and the control group (p<0.05), and the rest of the molecules also showed a trend of increasing in prostate patients.

[0174] Example 2. Screening of autoantibody combinations for prostate cancer detection

[0175] For the above tumor-associated autoantibody molecules, the present application comprehensively considers the positive detection rate and specificity of the candidate molecules in prostate cancer patients, while eliminating molecules with high overlapping positive detection (i.e. low individual positive contribution rate). Different molecules are selected from the aforementioned autoantibodies to form different autoantibody combinations, and corresponding detection reagents are used for detection, and the results are shown in Table 4 and Figures 12-13 .

[0176] Table 4. Sensitivity and specificity of autoantibody combinations as prostate cancer markers

[0177]

[0178] A preferred autoantibody combination is determined, which includes the following antibodies: Anti-COPB1, Anti-EIF3E, Anti-P53, Anti-CCDC110, Anti-BRCA1, Anti-MAGEA4, Anti-HNRPA1, and the sensitivity and specificity of the combination are 54.29% and 89.66%, respectively.

[0179] Example 3. Discrimination ability of the antibody combination of the present application for prostate cancer patients and healthy people

[0180] Using receiver operating characteristic curve (ROC) analysis, the preferred autoantibody combination of the present application has the ability to distinguish between healthy controls and prostate cancer patients in this study. This ROC curve is shown in Figure 2 When the Youden index reaches the maximum value, the sensitivity and specificity are 53.16% and 96.15%, respectively, and the area under the curve is 0.8598.

[0181] Example 4. Discrimination ability of the antibody combination of the present application for prostate cancer patients and benign disease patients

[0182] Using receiver operating characteristic curve (ROC) analysis, the preferred autoantibody combination of the present application has the ability to distinguish between benign disease and prostate cancer patients in this study. This ROC curve is shown in Figure 3As shown, when the Youden index reaches its maximum value, the sensitivity and specificity are 59.21% and 80.95%, respectively, and the area under the curve is 0.8111.

[0183] Example 5: The antibody combination of the present invention has better detection capability for prostate cancer patients with higher pathological grades (Gleason score).

[0184] This invention classifies the pathological grade of test patients according to the Gleason scoring system, classifying cases with a score of 8 or higher as high-grade prostate cancer and cases with a score of 7 or lower as low-grade prostate cancer. The invention analyzes the serum test data of different patient types to demonstrate the detection capability of the detection model for prostate cancer at different pathological grades.

[0185] like Figure 4 and 5 As shown, the autoantibody combination of the present invention exhibits a sensitivity of 57.14%, a specificity of 86.36%, and an area under the curve (AUC) of 0.7955 for low-grade prostate cancer; and a sensitivity of 62.50%, a specificity of 95.45%, and an AUC of 0.8523 for high-grade prostate cancer. This detection model demonstrates better detection capability for prostate cancer patients with higher pathological grades (i.e., higher malignancy).

[0186] Example 6. The antibody combination of the present invention has better detection capability for prostate cancer patients with low PSA levels.

[0187] This invention categorizes patients based on a PSA level of 10 ng / mL, classifying cases with PSA levels of 10 ng / mL or higher as high-PSA prostate cancer and cases with PSA levels less than 10 ng / mL as low-to-intermediate-PSA prostate cancer. Serum test data were analyzed for each patient type to assess the detection capability of the antibody combination against prostate cancer at different PSA levels. The invention found a 51.43% positive detection rate for the antibody combination in high-PSA patients and a 55.71% positive detection rate in low-PSA patients. This indicates that the antibody combination has higher sensitivity for prostate cancer patients with low PSA levels.

[0188] like Figure 6 and 7 As shown, at the maximum Youden index, the autoantibody combination of the present invention exhibits a sensitivity of 61.11%, a specificity of 91.30%, and an area under the curve of 0.8945 for low PSA prostate cancer; and a sensitivity of 53.13%, a specificity of 90.91%, and an area under the curve of 0.8196 for high PSA prostate cancer. This detection model demonstrates better detection capability for prostate cancer patients with low PSA levels.

[0189] Example 7. PSA alone as a prostate cancer marker has a weak ability to distinguish between benign and malignant prostate diseases

[0190] This example is directed to the subject population of the present application, and analyzes the ability of PSA alone as a prostate cancer marker to distinguish between healthy population or benign disease patients and prostate cancer patients.

[0191] When the threshold value of the present application is 10 ng / mL, the detection specificity of PSA is 100% and the sensitivity is 33.33% according to the detection results of PSA.

[0192] As shown in Figure 8 When the healthy population is used as the control, according to the roc analysis, the sensitivity and specificity of PSA at the maximum Youden index are 67.35% and 89.66%, respectively, and the area under the curve is 0.8033.

[0193] As shown in Figure 9 When the benign disease patients are used as the control, the sensitivity and specificity of PSA as a marker at the maximum Youden index are 40.00% and 93.48%, respectively, and the area under the curve is only 0.6253, which shows that PSA has a weak ability to distinguish between benign and malignant prostate diseases.

[0194] Example 8. The antibody combination of the present application has a higher cancer detection rate in patients with low and medium PSA prostate cancer

[0195] The present application divides the prostate cancer population into low PSA patients (4 ng / mL is the threshold value for clinically defining abnormal elevation of PSA), medium PSA patients (gray state of abnormal PSA value) and high PSA patients (10 ng / mL is usually used as the critical value for screening prostate cancer) with PSA values of 0-4 ng / mL, 4-10 ng / mL and above 10 ng / mL, respectively.

[0196] In the prostate cancer population, the antibody positive detection rate of low and medium PSA patients is higher, and the total antibody positive rate of these two parts is 55.71% [(14+25) / (31+39)], which shows good complementarity with PSA detection, and can have a better cancer detection rate in patients with low and medium PSA.

[0197] The preferred antibody combination positive rate of prostate cancer patients with different PSA levels is shown in Table 5.

[0198] Table 5

[0199]

[0200] A clinical study on prostate cancer screening conducted by Thompson IM et al. in the United States found that annual prostate cancer screening using PSA in the study population, at the end of the seven-year study, still had a 15.2% incidence of prostate cancer in the group that had never experienced an abnormal increase in PSA during the seven years. Among these patients, 14.9% had more malignant prostate cancer cases with a Gleason score of 7 or higher.

[0201] Therefore, it is evident that PSA has no predictive ability for a significant portion of prostate cancer cases, while the molecular combination of this invention shows significantly improved sensitivity in prostate cancer patients with low PSA compared to PSA alone. Incorporating this invention into prostate cancer screening could further reduce the harm caused by prostate cancer.

[0202] Example 9. The false positive rate of the antibody combination of the present invention is lower than that of PSA molecules.

[0203] In patients with benign prostatic hyperplasia, the false positive rate of the antibody combination was lower than that of PSA (with a threshold of 4 ng / mL).

[0204] Regarding the preferred autoantibodies, 9 out of 46 patients with benign prostatic hyperplasia tested positive for the antibody combination. For PSA, 29 out of the same 46 benign patients had PSA levels higher than the clinical threshold. Therefore, the false positive rate of the antibody combination was significantly lower than that of the clinically commonly used PSA molecule (19.57% vs. 63.04%), thus reducing the possibility of overdiagnosis.

[0205] Therefore, it can be seen that the preferred combination of autoantibodies of the present invention is more accurate than the currently clinically used PSA molecule as a biomarker for prostate cancer.

[0206] Table 6 shows the statistical results of false positive rates for autoantibody combinations and PSA (with a threshold of 4 ng / mL) in patients with benign diseases.

[0207] Table 6

[0208] False positive rate Preferred combinations of autoantibodies 19.57％ PSA (threshold at 4 ng / mL) 63.04％

[0209] Example 10: The antibody combination of the present invention, combined with PSA, further enhances the ability to distinguish between prostate cancer patients and healthy individuals.

[0210] As mentioned above, the antibody combination of the present application has good complementarity with PSA detection results, so we combined the two types of detection. Using receiver operating characteristic curve (ROC) analysis, the preferred antibody combination of the present application combined with PSA (i.e. the molecular combination of Anti-COPBl, Anti-EIF3E, Anti-P53, Anti-CCDC110, Anti-BRCA1, Anti-MAGEA4, Anti-HNRPA1 and PSA) had the ability to distinguish between healthy controls and prostate cancer patients in this study.

[0211] The positive detection rate of the combined molecular combination in prostate cancer patients was increased to 70.48%, which was significantly higher than that of the antibody combination alone. The ROC curve is shown in Figure 10 At the maximum Youden index, the sensitivity and specificity were 66.67% and 86.36%, respectively, and the area under the curve was 0.8793. Compared with the antibody combination alone, the performance of the combined detection model was further improved.

[0212] Example 11. The antibody combination of the present application combined with PSA also optimizes the ability to distinguish between prostate cancer patients and benign disease patients

[0213] Using receiver operating characteristic curve (ROC) analysis, the preferred antibody combination of the present application combined with PSA (i.e. the molecular combination of Anti-COPBl, Anti-EIF3E, Anti-P53, Anti-CCDC110, Anti-BRCA1, Anti-MAGEA4, Anti-HNRPA1 and PSA) had the ability to distinguish between benign disease and prostate cancer patients in this study.

[0214] The ROC curve is shown in Figure 11 At the maximum Youden index, the sensitivity and specificity were 65.22% and 80.95%, respectively, and the area under the curve was 0.8413. Compared with the antibody combination alone, the sensitivity and the area under the curve were significantly improved.

[0215] The ability of the antibody combination of the present application combined with PSA to distinguish between prostate cancer patients and healthy people, and between prostate cancer patients and benign disease patients, is shown in Table 7.

[0216] Table 7

[0217]

[0218] All documents referred to in the present application are incorporated herein by reference as if each were individually incorporated. In addition, it is to be understood that the application can be carried out by specifically different embodiments and that each disclosed embodiment can be implemented with or without the corresponding use of the other embodiments. Other embodiments will occur to readers of the disclosure and the appended claims.

Claims

1. Use of a diagnostic reagent for autoantibodies against a target antigen, characterized in that, A diagnostic reagent or kit for diagnosing the risk of prostate cancer is prepared, wherein the autoantibodies against target antigens include: (A1) Anti-COPB1; (A2) Anti-EIF3E; (A3) Anti-CCDC110; (A4) Anti-MAGEA4; (A5) Anti-HNRPA1; (B1) Anti-P53; and (B2) Anti-BRCA1. The diagnostic reagent of autoantibodies against target antigens is an antigen protein.

2. Use according to claim 1, characterized in that, The antigen protein includes: (C1) COPB1; (C2) EIF3E; (C3) CCDC110; (C4) MAGEA4; (C5) HNRPA1; (C9) P53; and (C10) BRCA1.

3. Use according to claim 2, characterized in that, The kit contains a detection reagent for detecting autoantibodies against target antigens, 4. A kit characterized in that, Wherein the autoantibodies against target antigens include: (A1) Anti-COPB1; (A2) Anti-EIF3E; (A3) Anti-CCDC110; (A4) Anti-MAGEA4; (A5) Anti-HNRPA1; (B1) Anti-P53; and (B2) Anti-BRCA1. The kit detects autoantibodies against target antigens through antigen-antibody reaction.

5. The kit of claim 4, wherein The kit is detected by the following methods: enzyme-linked immunosorbent assay (ELISA), protein / peptide segment chip detection, chemiluminescence, immunoblotting, microbead immunodetection, microfluidic immunization, or a combination thereof.

6. The kit of claim 4, wherein The kit also records the following detection methods, including steps:

7. The kit of claim 4, wherein (a) providing a detection sample; (b) detecting the level of autoantibodies against target antigens in the detection sample, denoted as Y1; And (c) comparing the level of autoantibodies against target antigens with the control reference value Y0; Wherein the autoantibodies against target antigens include: (A1) Anti-COPB1; (A2) Anti-EIF3E; (A3) Anti-CCDC110; (A4) Anti-MAGEA4; (A5) Anti-HNRPA1; (B1) Anti-P53; and (B2) Anti-BRCA1. If the detection result of autoantibodies against target antigens of the detection object meets the following conditions, it is suggested that the detection object has a high risk of prostate cancer: When the level of a certain autoantibody is higher than the reference value or standard value Y0, it is suggested that the detection object has a high risk of prostate cancer. The detection sample is selected from the group consisting of whole blood, serum, plasma, tissue, cells, interstitial fluid, cerebrospinal fluid, urine, or a combination thereof.

8. The kit of claim 7, wherein The device includes:

9. A prostate cancer risk determination device, comprising: (a) an input module for inputting autoantibody data against target antigens of a certain object; Wherein the autoantibodies against target antigens include: ​ (A1) Anti-COPB1; (A2) Anti-EIF3E; (A3) Anti-CCDC110; (A4) Anti-MAGEA4; (A5) Anti-HNRPA1; (B1) Anti-P53; and (B2) Anti-BRCA1; (b) a processing module: comparing the inputted autoantibody level Y1 with the control reference value Y0, so as to obtain a judgment result, wherein, when the comparison result meets a judgment condition, it is prompted that the subject has a high risk of prostate cancer; otherwise, it is prompted that the subject does not have a high risk of prostate cancer; (c) an output module, used for outputting the judgment result.

10. Use of a diagnostic reagent of an autoantibody-antigen combination, characterized in that, A diagnostic reagent or kit for jointly diagnosing the occurrence risk of prostate cancer is prepared; Wherein, the autoantibody-antigen combination comprises: autoantigen PSA; and The following autoantibodies: (A1) Anti-COPB1; (A2) Anti-EIF3E; (A3) Anti-CCDC110; (A4) Anti-MAGEA4; (A5) Anti-HNRPA1; (B1) Anti-P53; and (B2) Anti-BRCA1.

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