Two affinity immunoelectrophoretic methods for studying collagen interaction with von Willebrand factor antigen.

Two new immunoelectrophoretic methods are described for studying the interaction of collagen fibrils with von Willebrand factor antigen (vWF: Ag). In the first, the sample was electrophoresed through a collagen-agarose wedge into an antibody-agarose area, and immunoprecipitin lines was detected by staining. Different immunoprecipitin patterns were obtained with the vWF: Ag of normal plasma, commercial FVIII preparations, and von Willebrand disease (vWD) type IIa plasma as the result of collagen binding of vWF: Ag. In the other method, the sample was electrophoresed into agarose for preliminary separation of forms, followed by migration in the second dimension through a collagen spacer gel into an antibody-agarose area. This method demonstrated preferential binding of high molecular weight forms of vWF: Ag in normal plasma and slight binding of the lower molecular weight forms of antigen found in vWD type IIa plasma. The affinity wedge method is a convenient general method for finding quickly a useful concentration of affinity reagent.