Two novel monoclonal antibodies to VWFA3 inhibit VWF-collagen and VWF-platelet interactions.

BACKGROUND

The interaction of collagen-von Willebrand factor (VWF)-GPIb is essential for platelet adhesion, especially under high shear conditions. VWF, which acts as a bridge between platelets and exposed subendothelium, interacts with collagen through its A3 domain, which is a new target for the antithrombotic agent.

OBJECTIVE

To develop functional blockers that specifically inhibit VWF-dependent adhesion of platelets to collagen under high shear stress.

METHODS

To develop murine antihuman VWF A3 monoclonal antibodies (mAbs) by standard hybridoma technology, and characterize their abilities to block interactions between VWF A3 and collagen as well as platelet function.

RESULTS

Thirty anti-VWF-A3 mAbs were obtained. Among them, two mAbs, designated as SZ-123 and SZ-125, were found to inhibit VWF-collagen type III interaction. SZ-123 and SZ-125 inhibited the binding of purified human VWF (1.5 or 3 mug mL(-1)) to human placenta collagen type III (IC(50) = 0.07 +/- 0.02 and 0.15 +/- 0.03 mug mL(-1), respectively) or to calf skin collagen type III (IC(50) = 0.48 +/- 0.06 and 0.51 +/- 0.07 mug mL(-1), respectively) coated on plates. Under flow shear condition (1000 s(-1)), SZ-123 and SZ-125 inhibited platelet adhesion on human placenta collagen- or calf skin collagen-coated surfaces. Both mAbs also inhibited platelet aggregation induced by ristocetin, botrocetin or bovine plasma.

CONCLUSIONS

SZ-123 and SZ-125 inhibited VWF-collagen and VWF-platelet interactions.