Direct hapten-linked competitive inhibition enzyme-linked immunosorbent assay (CIELISA) for the detection of O-pinacolyl methylphosphonic acid
| Autor: | S. Merwyn, M. P. Kaushik, Ramarao Ghorpade, G. S. Agarwal, Manisha Sathe |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Ovalbumin
Soman Enzyme-Linked Immunosorbent Assay Binding Competitive Biochemistry Gas Chromatography-Mass Spectrometry Analytical Chemistry chemistry.chemical_compound Microtiter plate Non-competitive inhibition Electrochemistry medicine Animals Environmental Chemistry Methylphosphonic acid Spectroscopy Immunoassay Chromatography medicine.diagnostic_test Antibodies Monoclonal chemistry Glutaral Covalent bond Polystyrenes Female Rabbits Glutaraldehyde Haptens Hapten Conjugate |
| Zdroj: | The Analyst. 137:406-413 |
| ISSN: | 1364-5528 0003-2654 |
| DOI: | 10.1039/c1an15773f |
| Popis: | 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%. |
| Databáze: | OpenAIRE |
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