Direct hapten-linked competitive inhibition enzyme-linked immunosorbent assay (CIELISA) for the detection of O-pinacolyl methylphosphonic acid

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 t...

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Bibliographic Details
Published in:Analyst (London) 2012-01, Vol.137 (2), p.46-413
Main Authors: Sathe, Manisha, Ghorpade, R, Merwyn, S, Agarwal, G. S, Kaushik, M. P
Format: Article
Language:English
Subjects:
Analytical chemistry
Animals
Antibodies, Monoclonal - immunology
Antigens
Assaying
Binding, Competitive
Chemistry
Coating
Enzyme-Linked Immunosorbent Assay
Exact sciences and technology
Female
Gas Chromatography-Mass Spectrometry
Genetic algorithms
Glutaral - immunology
Haptens - chemistry
Haptens - immunology
Haptens - metabolism
Immunoassay
Miscellaneous
Networks
Ovalbumin - immunology
Plates
Polystyrene resins
Polystyrenes - chemistry
Rabbits
Soman - analogs & derivatives
Soman - analysis
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Summary: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 g mL 1 . The assay could detect PMPA even at the concentration of 0.006 g mL 1 . The mean recovery of standard PMPA (spiked in water) was found to be 83.7%. 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.
ISSN:0003-2654
1364-5528
DOI:10.1039/c1an15773f