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Thromb Haemost 1989; 61(01): 086-092
DOI: 10.1055/s-0038-1646532
Original Article
Schattauer GmbH Stuttgart

Functional Assays for Protein C Activity and Protein C Inhibitor Activity in Plasma

Margarethe Geiger
The Department of Immunology, Research Institute of Scripps Clinic, La Jolla, CA, USA
,
Teresa M White
The Department of Immunology, Research Institute of Scripps Clinic, La Jolla, CA, USA
,
John H Griffin
The Department of Immunology, Research Institute of Scripps Clinic, La Jolla, CA, USA
› Author Affiliations
Further Information

Publication History

Received 07 April 1988

Accepted after revision 20 September 1988

Publication Date:
24 July 2018 (online)

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Summary

An assay system for protein C (PC) activity and PC-inhibitor in plasma was developed. The assay was based on: (1) binding of PC to wells of a microtiter plate coated with a murine monoclonal anti-PC antibody (C3) that did not interfere with the activity or activation of PC; (2) activation of immobilized PC with Protac C; (3) incubation with or without a source of activated PC inhibitor; and (4) measurement of amidolytic activity using the substrate S-2366. The activity assay was specific for PC and sensitive to <1 μl of plasma or 4 ng PC. Inhibition of activated PC by plasma followed pseudo first order kinetics. Heparin caused a dose dependent increase in the inhibition rate with half maximal stimulation at approximately 3 U/ml and maximal stimulation at heparin concentrations ≥10 U/ml. This assay is suitable not only for determination of functional plasma levels of PC and PC inhibitor activities but also for kinetic studies of inhibition of activated PC in complex systems, such as plasma. Studies showed that urokinase interfered with the inhibition of APC by plasma inhibitor(s).

Key words

Protein C - Protein C inhibitor - Functional protein C assay - Functional protein C inhibitor assay
 

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