Development And Application For Monoclonal Antibody Of Ivermectin

Avermectins antibiotics are frequently used for killing the broad spectrum of parasite, mostly including Avermectin (AVM), Ivermectin (IVM) and Doramectin (DVM) etc. They are often useful for treating animal diseases in veterinary clinical and additives of animal feeds and drugs. However, illegal administration will lead to residues in edible tissues,and finally do latent harm to human health.Many countries and international organizations have established many methods to detect residue of IVM. Some methods are preciser and more sensitive ,but not to fit screening batch samples because of complicated method, wasting time, hypsi-cost and homo-technique .ELISA technique is gradually been applicating to detect residues of medicine as a method which is sensitive ,fast and powerful specificity.For explorating a method that can quickly detecting residue of ivermectin , this text composited man-made antigen, prepared anti-ivermectin monoclonal antibody ,finally, established ELISA method for detecting residues of ivermectin ,using monoclonal antibody technique and ELISA technique.Two man-made antigens for ivermectin were synthesized from succinic anhydride's method and EDC's method .used one of man-made antigens named IVM-BSA as a antigen for immune, a cell line (3D8) which excreted anti-ivermectin monoclonal antibody was gained by hybridoma's technique. The abdominal dropsy was got, and the potency was 1:32000; the saturation of protein in abdominal dropsy was 18.7mg/ml; the rate of crossing reaction with other antibacterial agents was lower than 0.1%.The CiELISA's method was made by the monoclonal antibody in the abdominal dropsy which was got by injecting hybridoma's cells named 3D8 into BALB/c mouse. The concrete method: the coating antigen was attenuated by 1:10000 and the monoclonal antibody in the abdominal dropsy was attenuated by 1.32000.The typical plot equations was y=37.203x-33.698(10～2000ng/ml).The correlate parameters of this equations as follow: LD50 was about 200ng/ml; the limit to detection was about 12ng/ml;the coefficient of differentiation in the same batch was 8.87% and the average coefficient of differentiation among other batches was 8.72%.The liver tissue and muscle tissue of swine were initially used for addition and reclamation test, and the sample was determined after pre-processing. Finally, the linear equation was y=32.612x-23.445(10～1000ng/),got by CiELISA. The recovery rate of sample was about 92%.