Western blot analysis of organophosphate exposure.
Division of Environmental Sciences, School of Public Health, Columbia University, New York, New York 1997 Mar; :1-56
A novel Western blot assay was developed which could detect acetylcholinesterase (AChE) from human erythrocytes. Isoelectric focusing (IEF) was used to separate at least five major isozymes of AChE. Inactivation of the catalytic activity with organophosphate pesticides did not change the isoelectric point of the isozymes, and therefore separation of native from organophosphate inhibited AChE did not occur. The degree of organophosphate inhibition did not influence the recognition of AChE by the monoclonal murine HR anti- AChEs. Native and organophosphate inactivated AChEs were not resolved by Western blotting that used alternative buffering systems. Treatment with organophosphates, diisopropylphosphorofluoridate (55914) and paraoxon (311455) produced aged AChE as evidence by resistance to reactivation with pralidoximine. The authors conclude that the charge difference between native and organophosphorus treated and aged intact AChEs is not sufficient for separation by IEF or electrophoresis. However, an internally controlled protein based assay would eliminate both interindividual and intraindividual variability, thus making it possible to assess AChE inhibition in the absence of preexposure values.
NIOSH-Grant; Grants-other; Pesticides; Blood-cells; Cholinesterase-inhibitors; Insecticide-poisoning; Organo-phosphorus-pesticides; Enzyme-activity
Environmental Sciences Columbia University 701 West 168Th Street New York, NY 10032-2704
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Other Occupational Concerns; Grants-other
Division of Environmental Sciences, School of Public Health, Columbia University, New York, New York
Columbia University New York, New York, New York