Pentachlorophenol monoclonal antibody hybridoma cell strain 2C3 and application thereof

A hybridoma cell line and monoclonal antibody technology, applied in the field of food safety immunological detection, can solve the problems of high cost, inability to realize rapid sample detection, time-consuming, etc., and achieve the effect of good detection sensitivity and specificity

Active Publication Date: 2016-04-06
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The method that detects pentachlorophenol at present mainly is gas chromatography (GC), liquid chromatography (HPLC), instrument methods such as spectrophotometry, derivative synchronous fluorescence method, thin-layer chromatography-spectrophotometry, gas chromatography-mass s

Method used

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  • Pentachlorophenol monoclonal antibody hybridoma cell strain 2C3 and application thereof
  • Pentachlorophenol monoclonal antibody hybridoma cell strain 2C3 and application thereof

Examples

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Embodiment 1

[0018] Embodiment 1: the preparation of hybridoma cell line 2C3

[0019] (1) Synthesis of complete antigen:

[0020] Add about 100mL of acetone solvent into a 250mL round-bottomed three-neck flask, under nitrogen protection, add 3g (12.15mmol) of tetrachlorohydrodiphenol and 10g (72.5mmol) of potassium carbonate, stir to dissolve as much as possible, and dissolve 1.58g (8.1mmol) Ethyl 4-bromobutyrate was dissolved in a small amount of acetone, added dropwise to the reaction solution, and started to reflux for about 12 hours. Thin-layer liquid chromatography (TLC) detected that the reaction was complete, and the acetone solvent was removed under reduced pressure. The reactants were dissolved in 50 mL of deionized water, and then 30 mL of ethyl acetate was added, fully shaken, allowed to stand and separated, and the water phase was retained. Add dilute hydrochloric acid solution to the water phase to adjust the pH to 3, extract the water phase with ethyl acetate 50mL×2, combin...

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Abstract

The invention discloses a pentachlorophenol monoclonal antibody hybridoma cell strain 2C3 and application thereof, and belongs to the field of food safety immunological detection. The pentachlorophenol monoclonal antibody hybridoma cell strain 2C3 is collected in the China General Microbiological Culture Collection Center (CGMCC) with a collection number CGMCC No.10871. A preparation method of the hybridoma cell strain 2C3 comprises the following steps: mixing a pentachlorophenol complete antigen and an equivalent Freund's adjuvant, and emulsifying the mixture, and immunizing a BALB/c mouse through back subcutaneous injection, wherein a complete Freund's adjuvant is used in a first immunizing process, and an incomplete Freund adjuvant is used afterwards; fusing spleen cells of the efficient and low-cost IC50 (Half Maximal Inhibitory Concentration) mouse with myeloma cells of the mouse through a PEG (Polyethylene Glycol) method, and performing indirect-competitive ELISA (Enzyme-Linked Immuno Sorbent Assay) screening and three times of sub-cloning to obtain a hybridoma cell strain 2C3. A monoclonal antibody secreted by the cell stain has high specificity and detection sensitivity (the IC50 value is 0.2mumg/L) specific to pentachlorophenol, and can be used for detecting residual pentachlorophenol in food safety.

Description

technical field [0001] The invention relates to a pentachlorophenol monoclonal antibody hybridoma cell line 2C3 and an application thereof, belonging to the field of food safety immunology detection. Background technique [0002] Pentachlorophenol, also known as pentachlorophenol, has a wide range of uses. As a pesticide, it is mainly its sodium salt. It can be used as a fungicide, preservative, herbicide, wood preservative, and partly as a solvent, heat exchange Waste from industrial production is released into the natural environment. Since the 1950s, my country has used pentachlorophenol and its sodium salt as the main pesticide to kill snails, the intermediate host of schistosomiasis. In 11 provinces, municipalities and autonomous regions in the middle and lower reaches of the Yangtze River in my country, a large area of ​​rice fields and ponds It has been continuously applied for decades, and its application was not gradually controlled until the 1980s. However, due to...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44G01N33/577
CPCC07K16/44C12N5/163G01N33/577
Inventor 匡华孙成胥传来徐丽广马伟刘丽强宋珊珊胡拥明
Owner JIANGNAN UNIV
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