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Antibodies against VIIIC (VIIICAg) were assayed using a modification of a two-site solid phase IRMA for factor VIII clotting antigen (VIIICAg). Anti-VIIICAg antibodies obtained from a multi- transfused haemophiliac were separated as IgG and labelled with I125. This was used to test plasma from patients with factor VIII inhibitor by competetive binding to common antigenic sites on immunoimobilised VIIIC. A haemophilic inhibitor assessed as 225u by the Bethesda method was used as standard. Results of inhibitor assay using the IRMA in 19 plasma samples from 15 severe haemophiliacs were similar to those obtained by the coagulation method. The increased sensitivity by IRMA of 0.01 u/ml enabled measurement of a haemophilic inhibitor undetectable by clotting assay. Anti VIIICAg activity was also detectable in plasmas from three individuals with acquired inhibitors against VIIIC. These plasmas which also had measurable residual VIIICAg gave dilution curves non-parallel to the standard haemophilic plasma curve. Measurement of haemophilic inhibitors using three IRMAs each employing different I125 labelled haemophilic anti VIIICAg antibodies showed that there was no difference in the sensitivity of the three assays but in some plasmas the results were discrepant indicating different specificities of the labelled antibodies.

Measurement Of Inhibitors To Procoagulant Factor VIII (VIIIC) By Immunoradiometric Assay (IRMA)