Preparation Of Monoclonal Antibody To OTC

Oxytetracycline (OTC) as the tetracycline class of veterinary drugs, itsantibacterial spectrum is broad, low prices, good efficacy advantages,OTC are widelyused in prevention and treatment of diseases;On the other hand, OTC also canimprove animals’ growth and development, so it often used as feed additives in animalfeed a large number of add, but if the feeder use of non-compliance with the provisionsof withdrawal period, there will be residues in animals.Once human consumption suchanimal foods, under the action of the food chain it will into the body and constantlyenriched, when OTC accumulated to a certain amount, it will cause harm to humanhealth. Therefore, It is necessary to strengthen the OTC drug residue detection. Atpresent, many countries in the world has a maximum residue limit (MRL) theprovisions of tetracyclines in animal foods, some countries even allowed thattetracycline cannot be detected in food.Traditional methods such as capillary electrophoresis and supercritical fluidchromatography, chromatography detection method, UV-Visible Spectrophotometryrequires large and expensive analytical instruments, is not suitable for field testing, andcost of testing is high, and detection process is complex, Detection time is very long, itis difficult to detect animal food in little time,and can not achieve field testing.Immunological detection technology is simple, short detection time, low cost, and it beable ti conduct on-site inspection. To establish the Immunoassay method of OTC, it isneed to Preparation the OTC specificity monoclonal antibody. The purpose of thisstudy is to preparation of OTC artificial antigen,Using monoclonal antibodytechnology to produce monoclonal antibodies specific for OTC, and provide a basis toestablish immunological rapid test.In this study, BSA and OVA were used as carrier protein,Using EDC method,successfully prepared the artificial antigen of OTC, UV spectrophotometry and SDS-PAGE, artificial antigen absorption peaks are offset or superimposedand, the colorreaction of artificial antigen identification were used to proved the preparation of artificial antigens is successful.In this study, the artificial antigens BSA-OTC was used to immune BALB/Cmice, obtained specific antibodies against OTC,50%PEG was used to fusion thespleen cells and myeloma cells, the two cells positive rate could reach100%. Throughthe cross-reaction experiments,it prove that the monoclonal antibodies which secretedby hybridoma cell lines has a high specificity.