Publication Date:
1997-02-01
Description:
We report the development of an enzyme-linked immunosorbent assay (ELISA) that is specific for factor VIIa (FVIIa). This assay uses a neoantigen specific capture antibody directed to the amino acid peptide sequence N terminal to the FVII cleavage activation site. The antibody exhibits ∼3,000-fold greater reactivity to FVIIa than FVII on a molar basis. Experiments using plasma with added (exogenous) human FVIIa gave quantitative recovery in the ELISA over a range of 0.20 to 3.2 ng/mL of FVIIa. The intra- and inter-assay coefficient of variation (CVs) of the ELISA are 4.5% and 9.8%, respectively. The ELISA shows excellent correlation (r = .99) with a functional assay (using recombinant soluble tissue factor) in detecting FVIIa added to plasma over the range 0.05 to 18.0 ng/mL. However, a major discrepancy exists between the two assays when normal endogenous plasma concentrations of FVIIa are measured. Using normal plasma (n = 14) the functional assay reported 3.10 ± 0.30 ng/mL (mean ± SE) whereas only 0.025 ± 0.010 ng/mL was detected in the same samples by the immunoassay. Patients (n = 43) presenting with acute coronary syndromes (myocardial infarction and unstable angina) exhibited elevations (P 〈 .05) in immunologically detected FVIIa, 0.093 ± 0.013 ng/mL (mean ± SE) compared to patient controls (n = 20) contemporaneously admitted with noncardiac chest pain, 0.048 ± 0.007 ng/mL (mean ± SE). These elevations in the acute coronary syndromes were accompanied by increased (P 〈 .05) and correlating prothrombin fragment F1 + 2 levels (Spearman correlation coefficient rs = .4, P 〈 .01), demonstrating that thrombin generation is certainly associated with, and may even be caused by, extrinsic pathway activation.
Print ISSN:
0006-4971
Electronic ISSN:
1528-0020
Topics:
Biology
,
Medicine
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