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Aristolochic acid A monoclonal antibody hybridoma cell strain and an application thereof

A hybridoma cell line, monoclonal antibody technology, applied in the direction of hybrid peptide, animal/human protein, serum albumin, etc., can solve the problems of low sensitivity, expensive equipment, and high requirements for the professional skills of experimental personnel, and achieve good specificity. The effect of high sensitivity and good detection sensitivity

Inactive Publication Date: 2018-12-14
JIANGNAN UNIV +1
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AI Technical Summary

Problems solved by technology

Thin-layer chromatography and ultraviolet spectrophotometry have low sensitivity; high-performance liquid chromatography (HPLC) has high sensitivity and accurate determination, and is currently the most commonly used method for determining aristolochic acid A, but the instruments and equipment are expensive and require professional staff high skill requirements

Method used

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  • Aristolochic acid A monoclonal antibody hybridoma cell strain and an application thereof
  • Aristolochic acid A monoclonal antibody hybridoma cell strain and an application thereof
  • Aristolochic acid A monoclonal antibody hybridoma cell strain and an application thereof

Examples

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Embodiment 1

[0023] Example 1 Preparation of hybridoma cell line SS0709

[0024] (1) Hapten: The chemical structural formula of aristolochic acid A contains an active group (-COOH), so it can be directly used as a hapten.

[0025]

[0026] (2) Preparation of complete antigen: Weigh 1.4 mg of aristolochic acid A, add 500 μL of DMF to dissolve, add EDC 2.5 mg and NHS 1.6 mg to it after fully dissolved, and stir at 30°C for 6 hours, as A solution. Weigh 10 mg of BSA, add 5 mL of CB to dissolve, and use it as solution B. Add liquid A droplet into liquid B, couple for 12 hours, dialyze the mixed liquid with 0.01M PBS solution, remove unreacted small molecule hapten, obtain the complete antigen AAs-EDC-BSA, and identify it by ultraviolet absorption scanning method;

[0027] (3) Mice immunization: After mixing and emulsifying the complete AAs antigen with an equal amount of Freund's adjuvant, BALB / c mice were immunized by subcutaneous injection at multiple points on the back of the neck (exce...

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Abstract

An aristolochic acid A monoclonal antibody hybridoma cell strain and an application thereof. The invention belongs to the field of food safety immunodetection. There is provided an aristolochic acid Amonoclonal antibody hybridoma cell strain SS0709, which is preserved in China General Microbiological Culture Collection Center and September 5th, 2017 and is assigned the accession number of CGMCC No.14688. An aristolochic acid A monoclonal antibody is secreted by the aristolochic acid A monoclonal antibody hybridoma cell strain SS0709 of which the accession number is CGMCC No.14688. The application of the aristolochic acid A monoclonal antibody is to analyze and detect the aristolochic acid A in traditional Chinese medicines. The monoclonal antibody secreted by the cell strain has excellentspecificity and detection sensitivity (IC50 being 4 ng / mL) to the aristolochic acid A, so that the monoclonal antibody can achieve the detection on the aristolochic acid A in the traditional Chinesemedicines. The cell strain supplies a raw material to the immunodetection on the aristolochic acid A in the traditional Chinese medicines and has practical application value.

Description

technical field [0001] The invention relates to an aristolochic acid A monoclonal antibody hybridoma cell line and an application thereof, belonging to the field of food safety immunoassay. Background technique [0002] Aristolochic acids (AAs for short), known as Aristolochic total acid, phagocytic acid or aketoxin, are a class of nitrophenanthrene carboxylic acids, and aristolochic acid compounds are aristolochic A class of nitrophenanthrene compounds unique to plants of the genus Aristolochia, including aristolochic acids A, B, C and D. In recent years, the nephrotoxicity of aristolochic acid has attracted widespread attention, and various countries have adopted control and restriction measures on traditional Chinese medicines containing aristolochic acid. On October 27, 2017, the list of carcinogens released by the International Agency for Research on Cancer of the World Health Organization was initially compiled for reference. Aristolochic acid and plants containing ar...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44C07K14/765G01N33/577C12R1/91
CPCC07K16/44C07K14/765C07K19/00G01N33/577
Inventor 胥传来李少珍匡华徐丽广马伟刘丽强吴晓玲宋珊珊胡拥明
Owner JIANGNAN UNIV
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