Tiamulin monoclonal antibody hybridoma cell line and application thereof

Abstract

The invention discloses a Tiamulin monoclonal antibody hybridoma cell line and application thereof, belonging to the field of food security immunity detection. The complete antigen of Tiamulin and equivalent Freund's adjuvant are mixed and emulsified completely, and then are injected into a BALB / c mice through back subcutaneous tissues so as to immunize the BALB / c mice; the first time of immunizing (100 mug / one mice) uses the complete Freund's adjuvant, immunity enhancement (50 mug / one mice) for multiple times uses the incomplete Freund's adjuvant, and finally, the complete antigen of the Tiamulin (20 mug / one mice, not containing adjuvant) is used for impacting immunity. Efficient but low-cost spleen cells of IC50 mice are fused with mice myeloma cells through a PEG method, and the hybridoma cell line is obtained through screening through an indirect cELISA and three times of subcloning. The monoclonal antibody secreted by the cell line has high specificity and detection flexibility (the value for IC50 is 0.73ng / mL) for the Tiamulin, and provides a raw material for immunity detection of residual Tiamulin in food, thereby having practical application value.

Description

technical field [0001] The invention relates to a tiamulin monoclonal antibody hybridoma cell line and an application thereof, belonging to the technical field of food safety immunology detection. Background technique [0002] Tiamulin (TML), also known as tiamulin, tiamulin, tiamulin, zhiyuanjing, tiamulin, is a special antibiotic for pleuromutilin, which is effective against mycoplasma and certain Some Gram-positive bacteria have good antibacterial activity and growth-promoting effect. They are rapidly absorbed in animals and widely distributed in the body, so they are widely used as veterinary drug antibiotics and feed additives. However, the residues of tiamulin in animals may pose a serious potential threat to human health. The European Union, the United States, and Japan have all made clear regulations on the maximum residue limit (MRL) of the antibiotic tiamulin in food of animal origin. my country's Ministry of Agriculture announced No. 235 in 2002 that the residue...

AI Technical Summary

Problems solved by technology

Although physical and chemical detection methods have high sensitivity, they require high professionalism of operators, take a long time, and are expensive. Immunological detection methods are characterized by fast sensitivity, simple operation, and high throughput, and have broad application prospects. It takes a long time to establish, once a good antibody is prepared, the establishment of a detection method will be very beneficial for the rapid detection of tiamulin

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