title: A quantitative solid-phase enzymeimmunoassay for 13,14-dihydro-15-keto- prostaglandin F2 alpha in plasma.
abstract: Enzymeimmunoassays (EIA) can be viable alternatives to radioimmunoassays (RIA). Indeed, from an environmental perspective, EIA are preferable to RIA. Therefore, the purpose of this project was to develop a quantitative EIA for 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) in bovine plasma. Acetylcholine esterase bound covalently to PGFM, rabbit anti-PGFM, mouse monoclonal anti-rabbit IgG, and PGFM were the principle reagents used for the EIA. Validation experiments indicated that: 1) PGFM standard curves, with doses ranging from 391 to 200,000 fg per microtiter well, were linear; 2) assay sensitivity averaged 391 fg per well; 3) for satisfactory results, PGFM had to be extracted from plasma; 4) content of PGFM in ethyl ether extracts of aliquots from serial dilutions of whole plasma with unknown amounts of PGFM and charcoal-stripped plasma supplemented with known amounts of PGFM did not deviate from parallelism with PGFM standard curves in buffer; 5) correlation between EIA and RIA measurements of PGFM in the same plasma samples was .95; 6) the regression of EIA data on RIA data was linear (Y = .93 x + 83.9; r2 = .91); 7) intra- and interassay coefficients of variation were 3.3 and 10.6%, respectively. The EIA developed in this project is a valid and reliable method for quantitating PGFM in extracts of bovine plasma.
authors.affiliation: Department of Animal Science, Virginia Polytechnic Institute and State University, Blacksburg 24061-0306.
authors.forename: R P
authors.initials: RP
authors.lastname: Del Vecchio
authors.forename: K M
authors.initials: KM
authors.lastname: Maxey
authors.forename: G S
authors.initials: GS
authors.lastname: Lewis
mesh_terms: D000818:Animals; D002021:Buffers; D002417:Cattle; D015237:Dinoprost / Q000031:analogs & derivatives* / Q000097:blood / Q000302:isolation & purification; D007124:Immunoenzyme Techniques; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D002021:Buffers; C008228:15-keto-13,14-dihydroprostaglandin F2alpha; D015237:Dinoprost