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Immunoradiometric assay (IRMA) for protein C was devised by using murine monoclonal antibodies against human protein C. The antibodies, specific for the activation peptide region of the heavy chain (HpC 9) or the light chain of human protein C (HpC 16), were used. HpC 9 was iodinated with a chlormine-T method and HpC 16 was immobilized to polyacrylamide beads (Immunobeads, Bio-Rad). The standard curves obtained either with purified human protein C or with pooled normal plasma with a known concentration of protein C were identical. The intraassay and interassay coefficients of variation for pooled normal plasma were 3.0% and 6.5% respectively. Twenty two normal individuals had a mean protein C level of 2.99±0/45mg/l (SD) and 9 warfarin-treated patients showed 1.26±0.24mg/l. Good correlation was obtained between this method and radioimmunoassay for human protein C in 22 normals, 9 warfarin-treated individuals and 3 patients with hereditary protein C deficiency (r=0.96, p<0.01).

Immunoradiometric assay for human protein C by using monoclonal antibodies against human protein C.