Preparation and identification of generic antigens and broad-spectrum monoclonal antibodies for detection of organophosphorus pesticides.
A simple, rapid, and high-sensitivity assay was developed to detect the multiresidue of organophosphorus pesticides (OPs) in the environment and food. Two separate generic haptens (Hapten A and B) with same O,O-dimethyl phosphorothioate group and aromatic ring and different spacer arms were synthesized and conjugated to bovine serum albumin (BSA) for immunogens and to ovalbumin (OVA) for coating antigens to study the effect of hapten and coating antigen heterology on immunoassay sensitivity. A broad-spectrum monoclonal antibody (MAb) was produced and a competitive ELISA developed using Hapten B-BSA as the immunogen and Hapten A-OVA as the coating antigen for the multiresidue determination of OPs, including parathionmethyl, fenitrothion, fenthion, chlorthion, and fenchlorphos. Several assay conditions, including organic solvent, pH, ionic strength, and incubation time, were studied sequentially to optimize the immunoassay. Using the optimal assay, 50% inhibition concentration values were estimated to be 34.5, 47.5, 79.8, 125.2, and 373.1 ng/mL for parathionmethyl, fenitrothion, fenthion, chlorthion, and fenchlorphos, respectively. The results indicated that the MAb showed specificity to all the above five OPs, and the assay could be developed for multiresidue determinations.
China Agricultural University, College of Food Science & Nutritional Engineering, 288 Mailbox, 17 Qinghua East Rd, Haidian District, Beijing 100083, People's Republic of China.
Panpan
P
Jiang
Jiguo
J
He
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000941:Antigens / Q000737:chemistry / Q000276:immunology; D002417:Cattle; D004785:Environmental Pollutants / Q000032:analysis / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005506:Food Contamination / Q000032:analysis; D006241:Haptens / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D057230:Limit of Detection; D051379:Mice; D008807:Mice, Inbred BALB C; D009946:Organothiophosphorus Compounds / Q000032:analysis* / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D010047:Ovalbumin / Q000737:chemistry / Q000276:immunology; D010573:Pesticide Residues / Q000032:analysis* / Q000276:immunology; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology
C570471:3-(4-(dimethoxy-O-phosphorothioyl)phenyl)propanoic acid; D000911:Antibodies, Monoclonal; D000941:Antigens; D004785:Environmental Pollutants; D006241:Haptens; C570472:O,O-dimethyl-O-(4-(carboxyl)phenyl)phosphorothiate; D009946:Organothiophosphorus Compounds; D010573:Pesticide Residues; D012710:Serum Albumin, Bovine; D010047:Ovalbumin