Development of a monoclonal antibody to factor VIII C2 domain and its functional study / 中华血液学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To develop a monoclonal antibody (mAb) directed to FVIII C2 domain and investigate its effect on FVIII activity.</p><p><b>METHODS</b>FVIII C2 protein was expressed in E. coli and purified. A murine antihuman FVIII C2 domain mAb SZ-132 was developed by standard hybridoma technology and characterized. In coagulation assays, different concentrations of SZ-132 were incubated with freshly collected pooled human plasma and the residual activity of FVIII and activated partial thromboplastin time (APTT) were determined. The effects of SZ-132 on rhFVIII binding to purified human vWF, phosphatidylserine (PS) and platelets were assessed by enzyme linked immunosorbent assays (ELISA).</p><p><b>RESULTS</b>SZ-132 could inhibit FVIII procoagulant activity in a dose-dependent manner within the concentrations of 0-25 microg/ml and the FVIII activity was completely inhibited on above 25 microg/ml. It could also prevent rhFVIII from binding to vWF, PS and platelets.</p><p><b>CONCLUSIONS</b>SZ-132 is a neutralizing mAb against FVIII C2 domain and can inhibit FVIII procoagulant activity by preventing FVIII from binding to vWF and PS.</p>