title: Enzyme-linked immunosorbent assay for detection of organophosphorylated butyrylcholinesterase: a biomarker of exposure to organophosphate agents.
abstract: A sandwich enzyme-linked immunosorbent assay (sELISA) has been developed for detection of organophosphorylated butyrylcholinesterase (OP-BChE), a potential biomarker for human exposure to organophosphate insecticides and nerve agents. A pair of antibodies specific to OP-BChE adduct were identified through systematic screening of several anti BChE antibodies (anti-BChE) and anti-phosphoserine antibodies (anti-P(ser)) from different sources. The selected anti-BChE (set as capture antibody) antibodies recognize both phosphorylated and nonphosphorylated BChE. These antibodies can therefore be used to capture both BChE and OP-BChE from the sample matrices. The anti-P(ser) (set as detecting antibody) was used to recognize the OP moiety of OP-BChE adducts. With the combination of the selected antibody pair, several key parameters (such as the concentration of anti-BChE and anti-P(ser), and the blocking agent) were optimized to enhance the sensitivity and selectivity of the sELISA. Under the optimal conditions, the sELISA has shown a wide linear range from 0.03 nM to 30 nM, with a detection limit of 0.03 nM. Furthermore, the sELISA was successfully applied to detect OP-BChE using in vitro biological samples such as rat plasma spiked with OP-BChE with excellent adduct recovery (z>99%). These results demonstrate that this novel approach holds great promise to develop an ELISA kit and offers a simple and cost-effective tool for screening/evaluating exposure to organophosphate insecticides and nerve agents.
authors.affiliation: Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture, Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, China.
authors.forename: Limin
authors.initials: L
authors.lastname: Wang
authors.forename: Dan
authors.initials: D
authors.lastname: Du
authors.forename: Donglai
authors.initials: D
authors.lastname: Lu
authors.forename: Chiann-Tso
authors.initials: CT
authors.lastname: Lin
authors.forename: Jordan N
authors.initials: JN
authors.lastname: Smith
authors.forename: Charles
authors.initials: C
authors.lastname: Timchalk
authors.forename: Fengquan
authors.initials: F
authors.lastname: Liu
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.forename: Yuehe
authors.initials: Y
authors.lastname: Lin
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D015415:Biomarkers / Q000097:blood; D002091:Butyrylcholinesterase / Q000097:blood* / Q000737:chemistry / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007306:Insecticides / Q000032:analysis*; D009943:Organophosphorus Compounds / Q000032:analysis*; D010766:Phosphorylation; D051381:Rats
chemical_list: D000906:Antibodies; D015415:Biomarkers; D007306:Insecticides; D009943:Organophosphorus Compounds; D002091:Butyrylcholinesterase