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A Lincosamide Universal Monoclonal Antibody Hybridoma Cell Line and Its Application

A monoclonal antibody, monoclonal technology, applied in instruments, peptides, immunoglobulins, etc., can solve the problems of expensive instruments, long time, and complicated operations

Active Publication Date: 2020-08-04
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the instrument detection method is accurate, but the instrument is expensive, the operation is complicated, the time is long, and the sample pretreatment is cumbersome.

Method used

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  • A Lincosamide Universal Monoclonal Antibody Hybridoma Cell Line and Its Application
  • A Lincosamide Universal Monoclonal Antibody Hybridoma Cell Line and Its Application
  • A Lincosamide Universal Monoclonal Antibody Hybridoma Cell Line and Its Application

Examples

Experimental program
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Embodiment 1

[0031] Example 1: Preparation of Lincosamides Universal Monoclonal Antibody

[0032] (1) Design of hapten: It is prepared by the condensation reaction of β-mercaptopropionic acid and clindamycin. For the specific synthesis route, see figure 1 . Dissolve 9.6g of clindamycin original drug in 20mL of dimethyl sulfoxide (DMSO), add 2.3g of KOH, slowly add β-mercaptopropionic acid solution (3.2g of β-mercaptopropionic acid dissolved in 10mL DMSO) dropwise while stirring, Heat the oil bath to 100°C and react for 2 h. After the product is naturally cooled to room temperature, add 50 mL of water, adjust the reaction solution to pH 3 with 6 mol / L HCl, and then extract with 50 mL of dichloromethane. The extract is dried over anhydrous Na2SO4, concentrated, Crystallization yielded yellow crystals, the target compound, which was identified by mass spectrometry.

[0033] (2) Preparation of the complete antigen: using clindamycin chloride-substituted derivatives, coupled with bovine serum...

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Abstract

A lincosamide general-purpose monoclonal antibody hybridoma cell line and application thereof belong to the technical field of food safety immunology detection. The present invention adopts clindamycin chloride substituted derivatives as hapten, couples hapten with bovine serum albumin (BSA) through activated ester method, obtains immune antigen, mixes with Freund's adjuvant evenly, and immunizes BALB / c mouse: Clindamycin was coupled to egg albumin (OVA) by the carbonyldiimidazole (CDI) method as a coating antigen for detection of mouse serum and cell supernatant. Splenocytes from immunized mice were fused with mouse myeloma cells by PEG method, screened by indirect ELISA and indirect competitive ELISA and subcloned three times to obtain a group-selective hybridoma cell line. The cell strain provided by the invention has good inhibition to clindamycin, lincomycin and pirithromycin, and can meet the demand for lincosamide polyresidue immunoassay products in the current market.

Description

technical field [0001] The invention relates to a hybridoma cell strain of lincosamide general monoclonal antibody and application thereof, belonging to the technical field of food safety immunology detection. Background technique [0002] Lincosamides are active against Gram-positive aerobic bacteria and Gram-positive or negative anaerobic bacteria. Lincomycin, clindamycin and piricomycin are commonly used in veterinary clinics. [0003] Residues of lincosamides in food of animal origin can cause renal dysfunction and increased drug resistance of Gram-positive bacteria. Both my country and the European Union have made relevant regulations on maximum residue limits in food of animal origin. Therefore, it is urgent to establish a A fast, sensitive and accurate detection method for lincosamide drug residues to ensure the safety of animal food. [0004] At present, the methods for detecting lincosamide drug residues mainly include high performance liquid chromatography (HPLC), ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/20C07K16/44G01N33/577C12R1/91
CPCG01N33/577C07K16/44G01N2430/00G01N33/9446C07K2317/92
Inventor 匡华郭玲玲胥传来徐丽广马伟刘丽强宋珊珊吴晓玲
Owner JIANGNAN UNIV