Development of monoclonal antibodies against pirimiphos-methyl and their application to IC-ELISA.
To detect the organophosphorus (OP) pesticide pirimiphos-methyl in grain samples, a monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was developed and optimized. By the active esters method, pirimiphos-methyl hapten A was conjugated to keyhole limpet hemocyanin to be used as the immunogen for the production of monoclonal antibodies, and pirimiphos-methyl hapten B was conjugated to ovalbumin to be used as coating antigen. By using the monoclonal antibody and the coating antigen, an IC-ELISA has been developed. Under the established optimized conditions, the IC-ELISA showed an IC50 of 4.2 ng/mL with a detection limit of 0.07 ng/mL. The IC-ELISA showed negligible cross-reactivity with other OP pesticides except with pirimiphos-ethyl. Recoveries of pirimiphos-methyl from spiked grain samples ranged from 83 to 96%.
Yang
Zheng-You
ZY
Division of Applied Life Science, Graduate School of Gyeongsan National University, Chinju, Gyeongnam 660-701, South Korea.
Kolosova
Anna Yu
AY
Shim
Won-Bo
WB
Chung
Duck-Hwa
DH
D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001667:Binding, Competitive; D002523:Edible Grain / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005506:Food Contamination / Q000032:analysis; D006241:Haptens / Q000276:immunology; D006433:Hemocyanins / Q000276:immunology; D001467:Hordeum / Q000737:chemistry; D007306:Insecticides / Q000032:analysis*; D009946:Organothiophosphorus Compounds / Q000032:analysis* / Q000276:immunology*; D012680:Sensitivity and Specificity; D014908:Triticum / Q000737:chemistry
D000911:Antibodies, Monoclonal; D006241:Haptens; D007306:Insecticides; D009946:Organothiophosphorus Compounds; C014153:pirimiphos methyl; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin