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Antihuman hemoglobin detection reagent strips and monoclone antibody contained therewith

A monoclonal antibody and hemoglobin technology, which is applied in the field of forensic medicine and immunological testing, can solve the problems such as the lack of specificity of anti-human hemoglobin antibody and the detection error.

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, some colloidal gold immunochromatographic detection reagent strips for detecting human hemoglobin at home and abroad, because the specificity of anti-human hemoglobin antibody does not meet the inspection standards of public security or justice, sometimes there are inspection errors, such as identifying animal blood as blood

Method used

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  • Antihuman hemoglobin detection reagent strips and monoclone antibody contained therewith
  • Antihuman hemoglobin detection reagent strips and monoclone antibody contained therewith
  • Antihuman hemoglobin detection reagent strips and monoclone antibody contained therewith

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Preparation of CGMCC 0970 hybridoma cell line B9 producing human hemoglobin monoclonal antibody

[0054] ① Immune mice: For each BALB / C mouse, inject 100 μg human hemoglobin (product of Sigma, USA) with complete Freund's adjuvant into limbs, subcutaneous multiple points and intraperitoneally, and then add incomplete adjuvant after 4 weeks. Intraperitoneal injection of the same dose as the same dose of the same dose, followed by tail vein injection 4 weeks later, and 3 days later, the mice were killed and the spleen was fused with SP2 / O cells according to the conventional method.

[0055] ② Screening and cloning: The fused cells were selected by HAT and observed with an inverted phase-contrast microscope, and live hybrid cells were selected. Coat the 96-well ELISA plate with 3 μg / ml human hemoglobin solution (note: set aside one well to be coated with 5% bovine serum albumin or 10% skimmed milk powder as a negative control), indirect ELISA method, which is abou...

Embodiment 2

[0060] Example 2 Preparation of anti-human hemoglobin colloidal gold immunochromatographic detection reagent strip

[0061] ① Anti-human hemoglobin monoclonal antibody ascites preparation: take out CGMCC 0970 hybridoma cell line B from liquid nitrogen 9 , recovered and cultivated by conventional methods. Inject an appropriate number of cultured cells into the abdominal 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.

[0062] ② Separation and purification of anti-human hemoglobin monoclonal antibody: by conventional n-octanoic acid plus ammonium sulfate method or affinity chromatography method, the latter is specifically: add 1 / 10 volume of 1.0mol / L Tris-HCl in mouse ascites, pH 8.0 buffer solution, then add saturated ammonium sulfate solution or solid ammonium sulfate to 50% saturation, after standing for 2 hours, centrifuge, precipitate with physiological PBS (20mmol / L disodium hydro...

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Abstract

A hybridoma cell strain B9 (CGMCC No.0970), its monoclonal antibody resistant to human haematoglobin, the reagent strip containing said monoclonal antibody 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 for the confirmation of human blood by forensic evidence. The antibody and the hybridoma cell line producing the monoclonal antibody, and the invention also relates to the preparation method of the detection reagent strip. Background technique [0002] In public security forensic work, the rapid 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 immunochromatographic detection reagent strips, similar immunological detection techniques 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 of them must be carried out...

Claims

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Application Information

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IPC IPC(8): C07K16/00C12N15/02G01N33/53G01N33/543G01N33/577
Inventor 李兆隆王俭张英兰徐秀兰王香菊常彩琴严红蒯应松陈艳
Owner 公安部第二研究所
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