1985 Volume 16 Issue 2 Pages 205-207
Monoclonal antibodies against von Willebrand factor (vWF) were produced by immunization of BALB/C mice with FVIII/vWF fraction and fusion of spleen cells with NS-1 murine myeloma cells. Antibody producing hybridomas were detected by an enzyme linked immunosorbent assay (ELISA). Five cloned monoclonal antibodies obtained in this method were all belonged to IgG1 class, and inhibited platelet retention by glass beads (Bowie et al.). The inhibitory action was most remarkable in 3B11. The antibody titer against vWF: Ag ranged from 1×105 to 1×107 by an ELISA. 3B11 inhibited ristocetin induced platelet aggregation and ristocetin cofactor up to 1×104. VIII protein and vWF protein were separately eluted from FVIII/vWF fraction by immunoadsorbent chromatography using monoclonal antibody. A new ELISA method was developed using each monoclonal antibody to quantitate vWF: Ag. The lower limit was 0.001u/ml. In order to detect vWF: Ag in tissues immunoperoxidase staining by avidin-biotin system using monoclonal antibody was performed. Positive staining was obtained in the endothelial cells of the neonatal umbilical cord, washed platelets and megakaryocytes.
