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Preparation method of anti-bisphenol A monoclonal antibody

A technology of monoclonal antibody and bisphenol, applied in the field of detection, can solve the problems of poor specificity of polyclonal antibody and difficult to meet the monitoring and analysis of bisphenol A, and achieve the effect of sensitive on-site monitoring and effective monitoring

Inactive Publication Date: 2010-10-20
GUANGDONG UNIV OF TECH
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Problems solved by technology

Due to the poor specificity and cross-reactivity of polyclonal antibodies, it is difficult to meet the needs of monitoring and analyzing trace amounts of bisphenol A. Therefore, if monoclonal cell lines secreting bisphenol A antibodies can be screened, a large number of monoclonal antibodies to bisphenol A can be produced. , the establishment of an immunoassay method for bisphenol A can more effectively monitor bisphenol A, and realize the rapid, specific and sensitive on-site detection of bisphenol A

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  • Preparation method of anti-bisphenol A monoclonal antibody
  • Preparation method of anti-bisphenol A monoclonal antibody
  • Preparation method of anti-bisphenol A monoclonal antibody

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Embodiment Construction

[0028] 1) Synthesis and identification of bisphenol A artificial immune antigen and coated antigen:

[0029] First, the structure of bisphenol A was modified, carboxyl groups were introduced, and the structurally modified bisphenol A was mixed with bovine serum albumin (BSA) and chicken eggs by carbodiimide and N-hydroxysuccinimide mixing method, respectively. Preparation of bisphenol A artificial immune antigen and coating antigen by coupling albumin (OVA). The artificial antigen was identified by ultraviolet spectrum and infrared spectrum, and the results showed that the synthetic artificial antigen had the characteristic absorption peaks of protein and bisphenol A in the ultraviolet spectrum, and the infrared spectrum of the artificial antigen showed the infrared characteristic absorption of bisphenol A and protein peak, the identification results indicated that the bisphenol A artificial antigen was synthesized successfully.

[0030] II) Preparation and screening of bisphen...

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Abstract

The invention discloses a preparation method of an anti-bisphenol A monoclonal antibody. The preparation method comprises the following steps of: combining bisphenol A and a macromolecule carrier protein to prepare an artificial immunity antigen and a coating antigen; preparing splenocyte suspension from a bisphenol A artificial immunity antigen immunity mouse; fusing the mouse splenocyte and mouse myeloma cells SP2 / 0 in the presence of polyethylene glycol; detecting by hybridoma technology and subcloning for many times to obtain positive hybridoma cells which can stably secrete bisphenol A antibody, wherein the secreted antibody belongs to the IgG1 subclass; and establishing a bisphenol A indirect competition ELISA (Enzyme-Linked Immunosorbent Assay) test by utilizing the monoclonal antibody secreted by cell lines with high antibodytiter, wherein the minimum limit of detection can reach 0.324ng / mL. The antibody has no cross reaction with compounds with benzene ring structures, such as benzene, phenol, tert-butylphenol, p-hydroxylphengl, o-hydroxybenzoic acid and the like, has strong specificity and can be used for immunity detection of bisphenol A.

Description

technical field [0001] The invention relates to a method for preparing a monoclonal antibody directed against the environmental hormone bisphenol A, and belongs to the technical field of detection. Background technique [0002] Bisphenol A (Biphenol A), referred to as BPA, is 2,2-bis(4-hydroxyphenyl)propane, crystalline or white scales, with a slight phenolic smell. Soluble in alkali solution, ethanol, acetone, acetic acid, ether and benzene, slightly soluble in carbon tetrachloride, almost insoluble in water. Bisphenol A is synthesized from phenol and acetone in an acidic medium, and is widely used in the production of beverage containers, tableware, baby bottles, pacifiers, and as one of the raw materials for tooth sealants and dental fillings. It is used in the manufacture of dyes, mechanical instruments It is widely used in industrial production and people's daily life, and its application is very common. [0003] Bisphenol A is an environmental estrogen substance, whi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/44C07K14/765C12N5/20G01N33/577
Inventor 赵肃清黄洪张磊张焜李明月郑杰方岩雄黄宝华
Owner GUANGDONG UNIV OF TECH
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