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Antihuman seminal specific protein P30 detection reagent strips and monoclone antibody therewith

A monoclonal antibody and specific protein technology, applied in biological testing, immunoglobulin, specific peptides, etc., can solve problems such as inspection errors, anti-human semen specific protein P30 antibody specificity not fully achieved, etc.

Inactive Publication Date: 2004-04-21
公安部第二研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, colloidal gold immunochromatographic detection reagent strips for detecting human semen spots at home and abroad, because the specificity of the anti-human semen-specific protein P30 antibody therein does not fully meet the inspection standards of public security or justice, sometimes there are inspection errors, and the Vaginal secretions have been identified as semen spots, and as we all know, police or judicial inspections do not allow false inspections, because it is related to a person's future or fate

Method used

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  • Antihuman seminal specific protein P30 detection reagent strips and monoclone antibody therewith
  • Antihuman seminal specific protein P30 detection reagent strips and monoclone antibody therewith
  • Antihuman seminal specific protein P30 detection reagent strips and monoclone antibody therewith

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Preparation of CGMCC 0971 hybridoma cell line A7 and CGMCC 0972 hybridoma cell line H3F3 producing anti-human semen-specific protein P30 monoclonal antibody:

[0054] According to Zhang Qi, Yan Hong, Wang Xiangju's "Chinese Journal of Forensic Medicine" 1990; 5(2): 72-76 pages "Establishment and preliminary application of hybridoma cell line secreting anti-human semen-specific protein P30 monoclonal antibody". After testing, a number of hybridoma cell lines that react only with human semen, purified human P30 fluid and human prostatic fluid were prepared, including CGMCC 0971 hybridoma cell line A7 and CGMCC 0972 hybridoma cell line H3F3. The antibodies produced are all mouse IgG1 subclass, and the titers are respectively, H3F3: 320 times of culture supernatant, ascites 1.28×10 5 times; A7: culture supernatant 32 times, ascites 1.0×10 4 times.

Embodiment 2

[0055] Example 2 Preparation of anti-human semen-specific protein P30 colloidal gold immunochromatography detection reagent strip

[0056] ①Ascites monoclonal antibody preparation: A7 and H3F3 cell lines were taken out from liquid nitrogen, revived and cultured by conventional methods. Inject an appropriate number of cultured A7 and H3F3 cultured cells into the peritoneal cavity of the mouse, kill the mouse 7-10 days later or when it is observed that it can be slaughtered, and collect the ascites of the mouse respectively.

[0057] ②Isolated and purified anti-human semen-specific protein P30 antibody: carried out by conventional n-octanoic acid plus ammonium sulfate method or affinity chromatography method, the latter specifically: add 1 / 10 volume of 1.0mol / L Tris-HCl to mouse ascites , pH 8.0 buffer solution, then add saturated ammonium sulfate solution or solid ammonium sulfate to 50% saturation, after standing for 2hr, centrifuge, precipitate with physiological PBS (20mmol / ...

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Abstract

A hybridoma cell strain A7 (CGMCC No.0971), a hybridoma cell strain H3F3 (CGMCC No.0972), their monoclonal antibodies resistant to human sperma specific protein P30, the reagent strip containing said monoclonal antibodies for the colloidal gold immunochromatographic test, and the preparing process of said reagent strip are disclosed. Its advantages are high sensitivity and high specificity.

Description

technical field [0001] The invention belongs to the technical field of forensic medicine and immunology testing, and can be directly used in the identification of human sperm spots by forensic evidence. Hybridoma cells for cloning antibodies, anti-human semen specific protein P30 colloidal gold immunochromatography detection reagent strip and preparation method of the reagent strip. Background technique [0002] In public security forensic work, the quick inspection and identification of case material evidence plays a decisive role in the timely detection of cases, the arrest of criminals or the release of criminal suspects. Before the appearance of colloidal gold immunochromatography detection reagent strips, similar techniques for immunological testing were carried out by methods such as immunoelectrophoresis, immunodiffusion, and enzyme-linked immunosorbent assay (ie, ELISA). The technical requirements of personnel are high, or the inspection takes a long time, and all o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/00C12N15/02G01N33/53G01N33/543G01N33/577
Inventor 李兆隆王俭徐秀兰张英兰张琦常彩琴严红王香菊孙云青陈东风
Owner 公安部第二研究所
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