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The nature of factor VIII related antigen (FVIIIRAG) was studied in endothelial cell cultures (EC), cryoprecipitate supernatant (cryo-S) and gel-filtered plasma fractions (GPF). Antigenic reactivity (AR) was assessed using the Laurell electroimmunoassay (LA) and a two-site immunoradiometric assay (IRMA) using a non-coagulation inhibitory rabbit antiserum raised against a factor VIII concentrate prepared by cryoprecipitation and gel filtration. Electrophoretic mobility (EM) of FVIIIRAG was assessed by two dimensional crossed immunoelectrophoresis (2DCIE), ristocetin co-factor activity (RiCoF) was estimated using fixed platelets. In normal plasma the results of LA, RiCoF assay and IRMA were in agreement. FVIIIRAG synthesised by EC showed increased EM and, in the IRMA decreased AR when compared to LA often with non-parallel dose response curves (DRC). This effect was not prevented by the presence of protease inhibitors in the culture medium. RiCoF activity was reduced compared with. LA and was related to the parallelism of the DRC in the IRMA. In cryo-S and the late eluting GPFs increased EM of FVIIIRAG was seen, and in these preparations also the DRC of the IRMA were not parallel to the normal plasma DRC. AR and RiCoF activity were disproportionately reduced when compared to the LA. It is concluded that the IRMA is sensitive to the presence of different forms of FVIIIRAG, particularly those which exhibit low levels of RiCoF activity and increased EM. some of these forms are present in normal plasma.

Different Forms of Factor VIII Related Antigens in Endothelial Cell Cultures, Cryoprecipitate Supernatant and Normal Plasma