EVIDENCE FOR AN ABNORMAL EXPRESSION OF THE COLLAGEN BINDING DOMAIN IN VON WILLEBRAND'S DISEASE TYPE II
A recently developed new series of monoclonal antibodies (MAbs) against the von Willebrand factor (vWf) included antibodies strongly inhibiting ( Mab vWf-41) and partly inhibiting ( Mab vWf-33) the collagen binding of vWf. We also characterized two Mabs with interacting properties against the ristocetin induced platelet aggregation (MAbs vWf-21 and vWf-39). These antibodies were conjugated with horse-radish peroxidase (HRP) and examined in different constructions forming two-site MAb ELISA's for plasma vWf:Ag and compared with polyclonal antibody ELISA. Symmetrical MAb-ELISA ( i.e. same Mab for extraction and detection) gave practical no dose-response in the standard assay, whereas any different combination of Mabs gave favourable dose-response relationships in sensitive ELISA's for vWf:Ag. Two different sandwiches were chosen using MAb vWf-33 and Mab vWf-41 at either side of the ELISA. These two assay models gave results of plasma from normal persons almost identical to those obtained with polyclonal antibody ELISA. Also in type I von Willebrand's disease these three assays performed very uniformly. In subtypes II plasma ( IIA: n=7; IIB: n=3, IIC: n=l, IID: n=i) . the assay using vWf-33 for coating and vWf-41-HRP for detection measured considerably lower than the polyclonal ELISA and the Mab-ELISA based on the opposite combination. We believe, that our results are indicative of a molecular defect in the collagen binding domain of vWf in subtype II plasma.