Preparation of a multi-hapten antigen and broad specificity polyclonal antibodies for a multiple pesticide immunoassay.
A multi-determinant artificial antigen was prepared by haptens of four pesticides (chlorpyrifos, triazophos, carbofuran and parathion methyl) conjugating to the carrier protein BSA in turn. Male New Zealand white rabbits were immunized with this multi-determinant immunogen to produce the polyclonal antibodies (PAbs), which can recognize the four pesticides. The PAbs displayed high level for each relative hapten-OVA conjugate, with the favorable titers of 4.49 x 10(4), 8.98 x 10(4), 2.24 x 10(4) and 1.86 x 10(4), for CHBu-OVA, THHe-OVA, BFNB-OVA and MP5-OVA, respectively. Characterization studies of the PcAbs showed that it has high affinity and specificity to the four relative pesticides. An indirect competitive ELISA was developed for multi-residue determination. The I(50) value for the four pesticides was 0.290, 0.065, 0.582 and 2.824 microg mL(-1), with the detection limit (I(10)) of 0.022, 0.005, 0.015 and 0.115 microg mL(-1) for carbofuran, triazophos, chlorpyrifos and parathion methyl, respectively. The linear rang was 0.016-2.000, 0.005-0.500, 0.010-2.000 and 0.063-5.000 microg mL(-1), respectively, for carbofuran, triazophos, chlorpyrifos and parathion methyl. Results indicated that, this study provided a new strategy to develop immunoassays through artificial antigen design for pesticides multi-residue determination.
Wang
Shu Ting
ST
Institute of Pesticide and Environmental Toxicology, Zhejiang University, Hangzhou 310029, China.
Gui
Wen Jun
WJ
Guo
Yi Rong
YR
Zhu
Guo Nian
GN
D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D000918:Antibody Specificity; D000941:Antigens / Q000737:chemistry*; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D006241:Haptens / Q000737:chemistry*; D007118:Immunoassay / Q000379:methods*; D008297:Male; D008956:Models, Chemical; D010047:Ovalbumin / Q000737:chemistry; D010575:Pesticides / Q000032:analysis* / Q000737:chemistry; D011817:Rabbits; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry
D000906:Antibodies; D000941:Antigens; D006241:Haptens; D010575:Pesticides; D012710:Serum Albumin, Bovine; D010047:Ovalbumin