title: Direct hapten-linked competitive inhibition enzyme-linked immunosorbent assay (CIELISA) for the detection of O-pinacolyl methylphosphonic acid.
abstract: Immunoassay detection of O-pinacolyl methylphosphonic acid (PMPA) employing direct coating of N-2-aminoethyl-O-pinacolyl methylphosphonate (hapten B) on microtiter plates is reported. Coating was achieved by covalently linking hapten B to a glutaraldehyde (GA) polymer network directly bound to the polystyrene (PS) surface of a standard 96-well microtiter plate. 4-(2-(O-Pinacolylmethylphosphoryl amino)ethyl amino)-4-oxobutanoic acid (hapten A)-ovalbumin (OVA) conjugate served as the coating antigen for comparison with direct hapten B-coated plates in the CIELISA format. The developed assay employing direct hapten B coated plates demonstrated enhanced sensitivity with the IC(50) value for PMPA being 0.027 μg mL(-1). The assay could detect PMPA even at the concentration of 0.006 μg mL(-1). The mean recovery of standard PMPA (spiked in water) was found to be 83.7%.
authors.affiliation: Process Technology Development Division, Defence R & D Establishment, Jhansi Road, Gwalior, 474002, India. manishasathe74@gmail.com
authors.forename: Manisha
authors.initials: M
authors.lastname: Sathe
authors.forename: R
authors.initials: R
authors.lastname: Ghorpade
authors.forename: S
authors.initials: S
authors.lastname: Merwyn
authors.forename: G S
authors.initials: GS
authors.lastname: Agarwal
authors.forename: M P
authors.initials: MP
authors.lastname: Kaushik
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry; D005976:Glutaral / Q000276:immunology; D006241:Haptens / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D007118:Immunoassay; D010047:Ovalbumin / Q000276:immunology; D011137:Polystyrenes / Q000737:chemistry*; D011817:Rabbits; D012999:Soman / Q000031:analogs & derivatives* / Q000032:analysis
chemical_list: D000911:Antibodies, Monoclonal; D006241:Haptens; D011137:Polystyrenes; C046573:pinacolyl methylphosphonic acid; D010047:Ovalbumin; D012999:Soman; D005976:Glutaral