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Use of human RGPR protein in preparing prostatic cancer diagnostic reagent kit

A diagnostic kit and technology for prostate cancer, applied in immunoglobulin, instruments, analytical materials, etc., can solve the problem of low diagnostic specificity of PSA diagnostic kits, and achieve the effects of sensitive testing, easy operation, and good reproducibility

Inactive Publication Date: 2007-08-01
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The invention provides the application of a human RGPR protein in the preparation of a test kit for detecting prostate cancer, so as to solve the problem of low diagnostic specificity of the current PSA diagnostic kit

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1, goat anti-human RGPR protein IgG antibody preparation

[0048] Take healthy male 50kg adult sheep and immunize with RGPR purified protein antigen. 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 RGPR protein IgG antibody. The titer was determined and frozen for later use.

Embodiment 2

[0049] Embodiment 2, preparation of rabbit anti-human RGPR protein IgG antibody

[0050] Healthy male adult rabbits (5kg) were taken and immunized with RGPR purified protein antigen. 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 RGPR protein IgG antibody. The titer was determined and frozen for future use. Embodiment 3, preparation of mouse anti-human RGPR protein IgG antibody

Embodiment 3

[0051] Healthy male adult guinea pigs were immunized with RGPR purified protein antigen. 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. Mouse anti-human serum was subjected to secondary precipitation with saturated ammonium sulfate and applied to an ion-exchange chromatography column to obtain mouse anti-human RGPR protein IgG antibody. The titer was determined and frozen for future use.

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Abstract

The present invention relates to the application of human RGPR protein in preparing prostatic cancer diagnosing reagent kit for early diagnosis of prostatic cancer and treating effect evaluation, and belongs to the field of biotechnology. The RGPR diagnosing reagent kit of the present invention is prepared through the following steps: antibody preparation, antibody marking, and antibody coating micro porous polyethylene plate. The present invention has the advantages of simple operation, high detecting sensitivity and high reproducibility.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to the application of human RGPR (regucalcingene promoter related protein) in the preparation of kits. It can be used for early diagnosis and evaluation of treatment effect of prostate cancer. Background technique [0002] At present, the only diagnostic kit for prostate cancer is the PSA diagnostic kit. PSA is a serine protease produced by prostate epithelial cells and secreted in serum, which has high prostate tissue specificity. PSA exists in three forms in the blood: the free molecular form (FPSA), the complex form with α12 antichymotrypsin (PSA-ACT) and the complex form with α22 macroglobulinase (PSA-α2M), of which only Very few exist in the form of FPSA. Under normal circumstances, the barrier composed of endothelial layer, basal cell layer and basement membrane maintains a low concentration of serum PSA, and only when tumors or other lesions destroy this ba...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/531C07K16/00
Inventor 潘玉琢赵雪俭张灵赵丹李扬计国义高洪文
Owner JILIN UNIV
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