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Kitasamycin monoclonal antibody hybridoma cell strain SML and application thereof

A hybridoma cell line, monoclonal antibody technology, applied in the preparation of hybrid peptides, sugar derivatives, animal/human proteins, etc. The effect of detection sensitivity

Inactive Publication Date: 2019-09-20
JIANGNAN UNIV
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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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  • Kitasamycin monoclonal antibody hybridoma cell strain SML and application thereof
  • Kitasamycin monoclonal antibody hybridoma cell strain SML and application thereof
  • Kitasamycin monoclonal antibody hybridoma cell strain SML and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Preparation of kitasarmycin monoclonal antibody

[0036] (1) Preparation of hapten: The hapten KIT-CMO was prepared by the CMO method.

[0037]The specific method is as follows: Weigh 200 mg of KIT into a round bottom flask, add 30 mg, 0.24 mmol of CMO, add 5 mL of methanol and 5 mL of 0.02 M NaHCO 3 The buffer solution was placed in a constant temperature magnetic stirring water bath at 60°C for 12 hours of reaction. After the reaction, the reaction liquid was blown dry with a nitrogen blower. Particulate matter (primarily removing NaHCO which provides an alkaline environment 3 particles), and the methanol phase was blown dry again to obtain white kitasamycin oximated derivatives (KIT-CMO). Store in a 4°C refrigerator for later use. Identification by liquid chromatography-mass spectrometry, as attached figure 1 shown.

[0038] (2) Preparation of complete antigen: The immunogen KIT-EDC-BSA was synthesized by the activated ester (EDC) method, and the coati...

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Abstract

The invention relates to a Kitasamycin monoclonal antibody hybridoma cell strain SML and an application thereof, and belongs to the field of food safety immune detection. The hybridoma cell line SML is deposited in the general microbiology center of the Council for the preservation and administration of microbial strains in China, with the preservation number of CGMCC No. 17392. The complete antigen of kitasamycin and equivalent freund's adjuvant are mixed and emulsified to immunize BALB / c rats. Complete freund's adjuvant is used for the first immunization, incomplete freund's adjuvant is used for several times to strengthen immunization, and finally, kitasamycin complete antigen is used for sprint immunization. Splenic cells of rats with high titer and low IC50 are fused with myeloma cells of rats by a PEG method, cultured and screened; then the cells are screened by indirect competitive enzyme-linked immunosorbent assay and subcloned three times to finally obtain a monoclonal antibody hybridoma cell strain. The monoclonal antibody secreted by the cell strain has good specificity and detection sensitivity to kitasamycin, can realize the detection of kitasamycin residues in aquatic products, and has practical application value.

Description

technical field [0001] The invention relates to a kitasarmycin monoclonal antibody hybridoma cell line SML and an application thereof, belonging to the field of food safety immunoassay. Background technique [0002] Kitamycin (KIT) is a multi-component macrolide antibiotic produced by Streptomyces. The antibacterial performance is similar to that of erythromycin, and it has a strong inhibitory effect on Gram-positive bacteria, such as Staphylococcus, Streptococcus pyogenes, Streptococcus viridans, Streptococcus pneumoniae, Bacillus tetanus, and Bacillus diphtheria. It is used to prevent bacterial infection and mycoplasma disease in livestock and poultry, as well as as a growth-promoting agent. KIT is a fat-soluble drug that will remain in the environment for a long time, causing water and soil pollution, and may enter the human body through the food chain, causing damage to human organs to a certain extent, and may even lead to the emergence of drug-resistant bacteria. Man...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/44C07K14/765C07H17/08C07H1/00C12R1/91
CPCC07H1/00C07H17/08C07K14/765C07K16/44C07K19/00
Inventor 胥传来郭玲玲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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