A colorimetric assay for plasma antithrombin III using a new synthetic peptide substrate (PS-915)

Clinica Chimica Acta; International Journal of Clinical Chemistry
K Asai, M Asai

Abstract

A colorimetric assay for plasma antithrombin III (ATIII) has been developed, using a new synthetic peptide substrate, PS-915 (H-D-Phe-Pro-Arg-CHA). PS-915 is freely water-soluble and its aqueous solution is stable. It is highly specific for thrombin and its Km value is about the same as that of S-2238. By enzyme hydrolysis, the new substrate liberates 3-carboxy-4-hydroxyaniline (CHA), which turns blue in color due to the complex formation with added alkaline-pentacyanoammine ferroate. The assay is a three-stage kinetic, one-point procedure, based on the method of antithrombin (heparin cofactor activity) determination. Since absorption measurements of the CHA are usually obtained at 700 nm, no blank correction is necessary even when hyperbilirubinemic, lipemic and/or hemolyzed plasma are used. The calibration curve for ATIII is linear over the range of 0-175% of normal and has a high degree of reproducibility. The results obtained by this method are well correlated with data obtained by the commonly used chromogenic assay or by radial immunodiffusion.

References

Apr 1, 1975·Thrombosis Research·O R OdegardU Abildgaard
Sep 15, 1977·Clinica Chimica Acta; International Journal of Clinical Chemistry·M F Scully, V V Kakkar
Jun 1, 1978·Thrombosis Research·L H KahléJ W ten Cate
Nov 1, 1974·Thrombosis Research·M BlombäckL Svendsen

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