Screening Of ScFv Antibody Against Pesticide Of Thiophosphates

This study first in organophosphate class general hapten H1, H3 on the basis of through active ester method to synthesize the artificial antigen, and verify the coupling of the artificial antigen, Determined the coupling is successful, and then build up resistance to organophosphAte gate in this experiment class Sc Fv primary antibody library, on the basis of the antibodantibodies for primary library for five rounds of enrichment competive.In the last two rounds of selection of 60 strains of monoclonal antibody libraries, and indirect ELISA detection,screscreening of the two strains can express high affinity Sc Fv antibody phage single strains. At the same time obtain soluble antibody.Organophosphorus pesticide varieties, high efficacy, uses broad,broadspectrum, there are various ways and effect, and different ways of effect can have very good effect.Is a very important branch of pesticides.With the development of science and technology, some of the risk-benefit ratio of organophosphorus pesticide are alternatives to traditional highly infficieninefficient organophosphorus pesticid e.Traditional pesticide residue detection relies mainly on the instrument analysis method, but need to be complicated, expensive equipment and professional operation personnel, together with sample preparation is trival time-consuming, high cost of testing, are difficult to meet the needs of high flux, rapid and on-lindetection.And biochemical detection method because of the specificity is strong, purposeful, and the characteristics of high sensitivity rapid simple accurate is increasingly becoming the emerging of organophosphorus pesticide residues detection method of the mainstream, this study used singly linked through genetic engineering antibody(Sc Fv) immune analysis methods of organophosphorus pesticide residues detection research.This study first in organophosphate class general hapten H1, H3 on the basis of through active ester method to synthesize the artificial antigen, and verify the coupling of the artificial antigen, Determined the coupling is successful, and then build up resistanceorganophosphate gate in this experiment class Sc Fv primary antibody library, on the basis of the antibodies for primary library for five rounds of enrichment competive.In the last two rounds of selection of 60 strains of monoclonal antibody libraries, and indirect ELISA detection, screening of the two strains can express high affinity Sc Fv antibody phage single strains. At the same time obtain soluble antibody.After preliminary optimization, get the end point of the two kinds of antibody titer of 1:32000, the linear regression equation of H1 B/B0 Log C = 62.58 + 62.58(r2 = 0.9894), half in the inhibition of antigen concentration 129.38 u g/m L, linear regression equation with H3 H3 B/B0 Log C = 67.32 + 149(r2 = 0.9808), half in the inhibition of antigen concentration 51.43 u g/m L.