Mouse Anti-VWF Recombinant Antibody (clone 82D6A3) (CAT#: PABZ-132)

Figure 1 Inhibition of baboon VWF binding to collagen by 82D6A3.

Different concentrations of 82D6A3 (20 μL) were incubated with undiluted baboon plasma (220 μL) for 30 minutes before addition to a collagen-coated plate. Bound VWF was detected with rabbit antihuman VWF antibodies. The data are the mean of 4 measurements obtained with the plasma of 2 different baboons

Wu, D., Vanhoorelbeke, K., Cauwenberghs, N., Meiring, M., Depraetere, H., Kotze, H. F., & Deckmyn, H. (2002). Inhibition of the von Willebrand (VWF)–collagen interaction by an antihuman VWF monoclonal antibody results in abolition of in vivo arterial platelet thrombus formation in baboons. Blood, 99(10), 3623-3628.

Figure 2 Effect of 82D6A3 on platelet adhesion under flow conditions.

Platelet deposition onto a human collagen type III-coated surface was studied as a function of the concentration of mAb 82D6A3 at a shear rate 2600 s⁻¹ (A) and as a function of the shear rate (B). Closed bars, no antibody; open bars, 2.3 μg/ml 82D6A3. Data are mean ± S.D. (n= 4, with blood from two different donors).

Vanhoorelbeke, K., Depraetere, H., Romijn, R. A., Huizinga, E. G., De Maeyer, M., & Deckmyn, H. (2003). A consensus tetrapeptide selected by phage display adopts the conformation of a dominant discontinuous epitope of a monoclonal anti-VWF antibody that inhibits the von Willebrand factor-collagen interaction. Journal of Biological Chemistry, 278(39), 37815-37821.

Figure 3 Immunofluorescence staining of collagen-bound VWF (left panel) with accessible GPIb binding domains (right panel).

A, B: Background signal without application of VWF on collagen III; C, D: VWF bound to collagen III. GPIb signals were found co-localized to the signals originating from anti-VWF immunofluorescence, clearly distinguishable especially on areas with higher VWF density indicated by arrows. E: Immunostaining of collagen-bound VWF after inhibition of VWF-collagen binding using mAb 82D6A3. Binding of VWF to collagen III under flow was nearly completely inhibited. Scale bar; 50 µm.

Fuchs, B., Budde, U., Schulz, A., Kessler, C. M., Fisseau, C., & Kannicht, C. (2010). Flow-based measurements of von Willebrand factor (VWF) function: binding to collagen and platelet adhesion under physiological shear rate. Thrombosis research, 125(3), 239-245.

Figure 4 von Willebrand factor (VWF) binding to horse tendon vs. human type III collagen.

Incubation in human type III (A) and Horm (B) collagen coated wells for 2 h at 37 °C of 3 μg/mL of VWF or H1786A-VWF in the absence or presence of the neutralizing antibody 82D6A3 (0– 10 μg/mL); binding is represented as specific Absorbance (A₄₉₀ nm, mean ± SEM from triplicate experiments) after subtraction of non-specific binding, measured in 1% BSA saturated wells without collagen (*P < 0.01 vs. VWF); incubation in human type III collagen coated wells of VWF, H1786A-VWF, ΔA1-VWF and ΔA3-VWF (1 μg/mL) as a function of the ionic strength, in NaCl and pH 7.3 adjusted 10 mM phosphate buffers (C); bound VWF was detected with HRP-conjugated polyclonal anti-VWF antibodies; representative tracings of association and dissociation of the indicated concentrations of H1786A-VWF during perfusion at 30 μg/mL over dextran-insolubilized pepsin-digested type III collagen (D).

Bonnefoy, A., Romijn, R. A., Vandervoort, P. A. H., Van Rompaey, I., Vermylen, J., & Hoylaerts, M. F. (2006). von Willebrand factor A1 domain can adequately substitute for A3 domain in recruitment of flowing platelets to collagen. Journal of Thrombosis and Haemostasis, 4(10), 2151-2161.