A quantitative solid-phase enzyme immunoassay for 13,14-dihydro-15-keto-prostaglandin F2 alpha in plasma.
1992
Del Vecchio R.P. | Maxey K.M. | Lewis G.S.
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 0.95; 6) the regression of EIA data on RIA data was linear (Y = 0.93X + 83.9; r2 = 0.91); 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.
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