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A pentachlorophenol monoclonal antibody hybridoma cell line 2c3 and its application

A hybridoma cell line and monoclonal antibody technology, which is applied in the field of food safety immunology detection, can solve the problems of high cost, inability to realize rapid detection of samples, cumbersome operation, etc., and achieve good detection sensitivity and specificity

Active Publication Date: 2018-10-02
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 spectrometry, but These methods have the disadvantages of cumbersome operation, time-consuming, and high cost, and cannot realize the rapid detection of a large number of samples. Therefore, it is of great significance to establish a fast and simple pentachlorophenol detection method.

Method used

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  • A pentachlorophenol monoclonal antibody hybridoma cell line 2c3 and its application
  • A pentachlorophenol monoclonal antibody hybridoma cell line 2c3 and its application

Examples

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Embodiment 1

[0018] Example 1: Preparation of hybridoma cell line 2C3

[0019] (1) Synthesis of complete antigen:

[0020] Add about 100 mL of acetone solvent to a 250 mL round-bottomed three-necked flask, under nitrogen protection, add 3 g (12.15 mmol) of tetrachloro-p-diphenol and 10 g (72.5 mmol) of potassium carbonate, stir to dissolve as much as possible, add 1.58 g (8.1 mmol) of Ethyl 4-bromobutyrate was dissolved in a small amount of acetone, added dropwise to the reaction solution, and heated to reflux for about 12 hours. By thin layer liquid chromatography (TLC) detection, the reaction was complete, and the acetone solvent was removed under reduced pressure. The reactant was dissolved in 50 mL of deionized water, and then 30 mL of ethyl acetate was added, fully shaken, and allowed to stand for stratification, and the aqueous phase was retained. Dilute hydrochloric acid solution was added to the aqueous phase to adjust the pH to 3, the aqueous phase was extracted with ethyl aceta...

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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 its application, belonging to the field of food safety immunology detection. Background technique [0002] Pentachlorophenol is also known as pentachlorophenol. Pentachlorophenol is widely used. As a pesticide, it is mainly its sodium salt. It can be used as a fungicide, preservative, herbicide, and can also be used as a wood preservative. Waste from industrial production is released into the natural environment. Since the 1950s, pentachlorophenol and its sodium salt have been used as pesticides to kill snails, the intermediate host of schistosomiasis, in my country. It was used continuously for several decades in the 1980s, and its application was gradually controlled in the 1980s. However, due to the fact that schistosomiasis has not been effectively eradicated in some areas of southern my country, the epidemic situation has developed in some areas in recent...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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