Antibody Against A Family Of Pyrethroid Pestcides Was Prepared And The Research On Immunoassay Technology For Pestcides Cyhalothrin

Immunological analysis technique is an important method for detection of the pesticide residue in the world. In this work, polycolonal antibodies against a familiy of the pyrethrins widely used in our country were prepared. The configuration analogy with pyrethroid was synthesized from the intermediate of 3-Phenoxybenzoic acid as the major materials, the immunogen antigen conjugates were prepared by linking the PBA to big protein molecular of bovine serum albumin(BSA) with NHS/DCC, the coating antigen conjugates were prepared by linking the PBA to big protein molecular of ovalbumin(OVA) with mixed anhydride method(MA), the coupling reaction was confirmed a success by UV-visible light spectrum scanning.Antibody #0631 #0629 #0977 were prepared against the coupled PBA-BSA, and their antiserum gradient riched 2.0 105,8.2 105, 4.1 105 respectively; the cross-reaction rate to the other Pyrethroid pestcides ,the polycolonal can recognize some kinds of Pyrethroid, such as cyhalothrin, permethrin, cypermethrin, fenvalerate, cyfluthrin. Cyhalothrin eventually fixed for the competitive indirect enzyme-linked immunoassay method . The result as follows: An assay curve Y was formed by. the linking rate logit( B/Bo )and X, the logarithm of the r-Cyhalothrin concentration ( g/mL ) as the two influencing factors, in the range of 1-10000g/mL ,the two factor represent a apparent linear relation, the equation is as follows: Y=-0.8129X+2.3439, the R2 reaches0.9965 ,I50=0.76mg/mL, the inter-group CV reaches 4.7%,ex-group CV reaches 4-7.1 %.The conditions for competitive ELISA: coating of PBA-OVA is at 6g/mL the antibody working concentration is 6000 times dilution of the antiserum, the block of OVA is l%,blocking time is 2h. In the reaction system PBS, the ion concentration is 0.2~0.3mol/L, methanol is at 10%, the optional pH is 7 -9, reaction time is lh,TMB color time is 10min.