Anti-natamycin monoclonal antibody hybridoma cell strain and application thereof

Abstract

The invention discloses an anti-natamycin monoclonal antibody hybridoma cell strain and application thereof, and belongs to the technical field of food safety immunological detection. The scheme adopted by the invention is as follows: mixing and emulsifying natamycin complete antigen with an equal amount of complete freund's adjuvant, and then immunizing BALB / c mouse through subcutaneous injection on the back of the mouse, wherein the first immunization uses the complete freund's adjuvant, and subsequent immunization uses an incomplete freund's adjuvant; fusing splenocytes of the mouse with a high titre and a low IC50 value and myeloma cells of the mouse through a PEG method; performing indirect competitive ELISA screening and three times of subcloning to obtain a hybridoma (monoclonal) cell strain. Monoclonal antibodies secreted by the hybridoma (monoclonal) cell strain has relatively good specificity and detection sensitivity (the value of IC50 is 2.2 microgram / L) for natamycin, and can be used for detecting residual natamycin in food safety.

Description

technical field [0001] The invention discloses an anti-natamycin monoclonal antibody hybridoma cell line and application thereof, belonging to the technical field of food safety immunology detection, and relates to a hybridoma cell line M and an anti-natamycin monoclonal antibody produced therefrom. Background technique [0002] Natamycin is a natural, broad-spectrum, efficient and safe inhibitor of filamentous fungi such as yeast and mold, belonging to polyene macrolides. It not only inhibits fungi, but also prevents the production of mycotoxins. It can be widely used in food preservation and antifungal treatment. Natamycin has no inhibitory effect on bacteria, so it does not affect the natural ripening process of yogurt, cheese, raw ham, dry sausage. In June 1982, the U.S. FDA officially approved natamycin as a food preservative. In 1996, Natamycin was officially approved by China Food Additives Standardization Chemical Technical Committee as a food preservative. [00...

AI Technical Summary

Problems solved by technology

The biological potency determination method has high requirements on the experimental environment factors (temperature, etc.), material factors (culture medium, etc.), operating technology and other conditions, and the operation process is relatively cumbersome; the microplate bioassay method is suitable for large quantities of fermentation broth or samples detection; UV spectrophotometry is generally used as a routine control for natamycin detection, if it is used for quantitative analysis, the requirements for experimental conditions are relatively high; elemental analysis, colorimetric analysis and thin layer chromatography are mainly used in the qualitative analysis of natamycin; high performance liquid chromatography (HPLC) focuses on the detection and analysis of natamycin residues in high-purity natamycin products and food, but the sample pretreatment of this method is loaded down with trivial details, Time-consuming, expensive instruments, requiring professionals to operate, unable to achieve simultaneous detection of a large number of samples

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