The assay system for prothrombin: a two-stage assay using ecarin as the prothrombin activator and the enzyme-linked immunosorbent assays

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Title
The assay system for prothrombin: a two-stage assay using ecarin as the prothrombin activator and the enzyme-linked immunosorbent assays
Author(s)
Young Do Yoo; Moon Hi Han; Ji Young Kim
Bibliographic Citation
Korean Biochemical Journal, vol. 21, no. 2, pp. 169-175
Publication Year
1988
Abstract
In a two-stage assay of prothrombin, prothrombin was activated to thrombin by ecarin, the prothrombin activator from the venom of Echis carinatus, in the absence of calcium ions and phospholipid. The activation was almost complete within 30 minutes and the thrombin generated was not inhibited by the ecarin activation mixtures for a incubation time up to at least 20 hours, while it was inhibited by the absorbed plasma- serum mixture after 9-minute incubation. Two different substrates, fibrinogen and a synthetic chromogenic substrate, were used for assays of the thrombin activity and compared in their sensitivies in detection of prothrombin. The standard curve was linear between 2 ${\mu}g/ml$ of prothrombin concentration in the clotting assay, and between 0.2 ${\mu}g/ml$ and 16 ${\mu}g/ml$ in the chromogenic assay. In addition we developed the enzyme-linked-immunosorbent assays for the direct measurement of prothrombin. The minimum detectable concentrations of prothrombin by the indirect ELISA and the sandwich ELISA were 40 ng/ml and 3 ng/ml, respectively. The prothrombin level in bovine plasma was determined to be 224 ng/ml by the sandwich ELISA. The reliability of the sandwich ELISA was demonstrated by the within runs and between runs tests, which were 9.3% and 15.8%, respectively.
ISSN
0368-4881
Publisher
Korea Soc-Assoc-Inst
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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