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Western blot analysis of organophosphate exposure.

Authors
Jeffrey-AM; Grassman-J
Source
Division of Environmental Sciences, School of Public Health, Columbia University, New York, New York 1997 Mar; :1-56
Link
NIOSHTIC No.
00238177
Abstract
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.
Keywords
NIOSH-Grant; Grants-other; Pesticides; Blood-cells; Cholinesterase-inhibitors; Insecticide-poisoning; Organo-phosphorus-pesticides; Enzyme-activity
Contact
Environmental Sciences Columbia University 701 West 168Th Street New York, NY 10032-2704
CAS No.
55-91-4; 311-45-5
Publication Date
19970314
Document Type
Final Grant Report
Funding Amount
41003
Funding Type
Grant
Fiscal Year
1997
NTIS Accession No.
PB97-207112
NTIS Price
A05
Identifying No.
Grant-Number-R03-OH-03256
NIOSH Division
OEP
Priority Area
Other Occupational Concerns; Grants-other
Source Name
Division of Environmental Sciences, School of Public Health, Columbia University, New York, New York
State
NY
Performing Organization
Columbia University New York, New York, New York
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