Applications of polypeptides PSA2 for producing prostate gland carcinoma diagnose kit

Abstract

The invention relates to an application of human polypeptide PSA2 in a preparation of a prostate gland cancer diagnosis kit, which belongs to the biotechnology field, and the invention can be used for an early diagnosis of prostate gland cancer and an evaluation of treatment effect. The invention discloses a preparation of human recombinant polypeptide PSA2 diagnostic reagent kit, and the preparation comprises an antibody preparation, a labeled antibody and a preparation of peridium antibody polystyrene micro-plates. The invention has the advantages that the operation of the kit is simple, the test is sensitive and the reappearance is good. According to a plurality of experiments of the detecting of prostate gland cancer, and each milliliter human recombinant polypeptide PSA2 is counted as one micro-gramme, the detecting reference range of the prostate gland cancer is: when each milliliter human recombinant polypeptide PSA2 is lower than 5nanogram, the prostate gland cancer is negative; when each milliliter human recombinant polypeptide PSA2 is higher than 5nanogram, the prostate gland cancer is positive.

Description

technical field

[0001] The invention belongs to the field of biotechnology, and the patent mainly relates to the application of PSA2 in the preparation of protein and gene diagnosis kits. It can be used for early diagnosis of prostate cancer and monitoring of treatment effect. Background technique

[0002] Since the use of prostate specific antigen (PSA) in population screening, the diagnosis of prostate cancer has undergone epoch-making changes. As early as 1983, the United States first used PSA to screen prostate cancer in the population, and it took 20 years to achieve early diagnosis and treatment. However, PSA that only has prostate tissue specificity still has the following disadvantages: ①Only 25-30% of the people with PSA>4.0ng / ml in the group census are diagnosed with prostate cancer, and the invalid biopsy rate is as high as 70%; ②PSA has no effect on prostate cancer. Special types of cancer other than adenocarcinoma, such as signet ring cell carcinoma and squ...

Examples

Embodiment 1

[0089] Embodiment 1, goat anti-human recombinant polypeptide PSA2 IgG antibody preparation

[0090] Take healthy male 50kg adult sheep and immunize with human recombinant polypeptide PSA2 purified protein antigen / synthetic polypeptide. For the first time, the antigen was mixed with BCG and Freund's adjuvant, injected into sheep neck lymph nodes and subcutaneous muscle tissue, once every two weeks, a total of 4-5 times of immunization, the amount of antigen 5mg / time. Blood was collected two weeks after the last immunization, and the goat anti-human serum was subjected to secondary precipitation with saturated ammonium sulfate and then applied to an ion-exchange chromatography column to obtain goat anti-human recombinant polypeptide PSA2 IgG antibody, which was tested for titer and frozen for later use.

Embodiment 2

[0091] Example 2, preparation of rabbit anti-human recombinant polypeptide PSA2IgG antibody

[0092] Take healthy male adult rabbits (5kg) and immunize them with human recombinant polypeptide PSA2 purified protein antigen / synthetic polypeptide. For the first time, the antigen was mixed with BCG and Freund's adjuvant, and injected into the subcutaneous muscle tissue of rabbits, once every two weeks, a total of 4 times of immunization, the amount of antigen was 0.5mg / time. Blood was collected two weeks after the last immunization. Goat anti-human serum was subjected to secondary precipitation with saturated ammonium sulfate and applied to an ion-exchange chromatography column to obtain rabbit anti-human human recombinant polypeptide PSA2 IgG antibody. The titer was determined and frozen for future use.

Embodiment 3

[0093] Embodiment 3, preparation of mouse anti-human recombinant polypeptide PSA2 IgG antibody

[0094] Take healthy male adult guinea pigs and immunize them with human recombinant polypeptide PSA2 purified protein antigen / synthetic polypeptide. For the first time, the antigen was mixed with BCG and Freund's adjuvant, injected into the subcutaneous muscle tissue of guinea pigs, once every two weeks, a total of 4 times of immunization, the amount of antigen was 0.1mg / time. Blood was collected two weeks after the last immunization. The mouse anti-human serum was subjected to secondary precipitation with saturated ammonium sulfate and applied to an ion-exchange chromatography column to obtain the mouse anti-human recombinant polypeptide PSA2 IgG antibody. The titer was determined and frozen for future use.

technical field

[0001] The invention belongs to the field of biotechnology, and the patent mainly relates to the application of PSA2 in the preparation of protein and gene diagnosis kits. It can be used for early diagnosis of prostate cancer and monitoring of treatment effect. Background technique

[0002] Since the use of prostate specific antigen (PSA) in population screening, the diagnosis of prostate cancer has undergone epoch-making changes. As early as 1983, the United States first used PSA to screen prostate cancer in the population, and it took 20 years to achieve early diagnosis and treatment. However, PSA that only has prostate tissue specificity still has the following disadvantages: ①Only 25-30% of the people with PSA>4.0ng / ml in the group census are diagnosed with prostate cancer, and the invalid biopsy rate is as high as 70%; ②PSA has no effect on prostate cancer. Special types of cancer other than adenocarcinoma, such as signet ring cell carcinoma and squ...

Examples

Embodiment 1

[0089] Embodiment 1, goat anti-human recombinant polypeptide PSA2 IgG antibody preparation

[0090] Take healthy male 50kg adult sheep and immunize with human recombinant polypeptide PSA2 purified protein antigen / synthetic polypeptide. For the first time, the antigen was mixed with BCG and Freund's adjuvant, injected into sheep neck lymph nodes and subcutaneous muscle tissue, once every two weeks, a total of 4-5 times of immunization, the amount of antigen 5mg / time. Blood was collected two weeks after the last immunization, and the goat anti-human serum was subjected to secondary precipitation with saturated ammonium sulfate and then applied to an ion-exchange chromatography column to obtain goat anti-human recombinant polypeptide PSA2 IgG antibody, which was tested for titer and frozen for later use.

Embodiment 2

[0091] Example 2, preparation of rabbit anti-human recombinant polypeptide PSA2IgG antibody

[0092] Take healthy male adult rabbits (5kg) and immunize them with human recombinant polypeptide PSA2 purified protein antigen / synthetic polypeptide. For the first time, the antigen was mixed with BCG and Freund's adjuvant, and injected into the subcutaneous muscle tissue of rabbits, once every two weeks, a total of 4 times of immunization, the amount of antigen was 0.5mg / time. Blood was collected two weeks after the last immunization. Goat anti-human serum was subjected to secondary precipitation with saturated ammonium sulfate and applied to an ion-exchange chromatography column to obtain rabbit anti-human human recombinant polypeptide PSA2 IgG antibody. The titer was determined and frozen for future use.

Embodiment 3

[0093] Embodiment 3, preparation of mouse anti-human recombinant polypeptide PSA2 IgG antibody

[0094] Take healthy male adult guinea pigs and immunize them with human recombinant polypeptide PSA2 purified protein antigen / synthetic polypeptide. For the first time, the antigen was mixed with BCG and Freund's adjuvant, injected into the subcutaneous muscle tissue of guinea pigs, once every two weeks, a total of 4 times of immunization, the amount of antigen was 0.1mg / time. Blood was collected two weeks after the last immunization. The mouse anti-human serum was subjected to secondary precipitation with saturated ammonium sulfate and applied to an ion-exchange chromatography column to obtain the mouse anti-human recombinant polypeptide PSA2 IgG antibody. The titer was determined and frozen for future use.

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