Improved Characterization of Plasma von Willebrand Factor Heterogeneity when Using 2.5% Agarose Gel Electrophoresis

Summary

Discontinuous sodium dodecyl sulfate electrophoresis in large pore gels, followed by overlay with radiolabelled anti-von Willebrand factor (vWF) antibodies and by autoradiography, permits to analyze the multimeric structure of vWF. The aim of this study was to improve experimental conditions of this technique to satisfactorily resolve the minor forms of plasma vWF while still separating its high, intermediate, and low molecular weight predominant multimers. By using a 2.5% mixture of two selected agaroses, a single electrophoretic analysis of plasma clearly reveals the extreme complexity of the molecular forms of circulating vWF: each multimeric unit of plasma vWF may be separated into five bands, the central one being predominant. The multimeric distribution and “quintuplet” pattern obtained in the electrophoretic system described here permit a convenient classification of the different subtypes of von Willebrand’s disease.

Key words

von Willebrand factor - von Willebrand’s disease - SDS - agarose electrophoresis

• References

• 1 Hoyer LW, Shainoff JR. Factor VUI-related protein circulates in normal human plasma as high molecular weight multimers. Blood 1980; 55: 1056-1059
• 2 Ruggeri ZM, Zimmerman TS. Variant von Willebrand’s disease. Characterization of two subtypes by analysis of multimeric composition of factor VUI/von Willebrand factor in plasma and platelets. J Clin Invest 1980; 65: 1318-1325
• 3 Ruggeri ZM, Zimmerman TS. The complex multimeric composition of factor VIII/von Willebrand factor. Blood 1981; 57: 1140-1143
• 4 Kinoshita S, Harrison J, Lazerson J, Abildgaard CF. A new variant of dominant type II von Willebrand’s disease with aberrant multimeric pattern of factor VIII-related antigen (type IID). Blood 1984; 63: 1369-1371
• 5 Tuddenham EG D. The varieties of von Willebrand’s disease. Clin Lab Haemat 1984; 6: 307-323
• 6 Ruggeri ZM, Nilsson IM, Lombardi R, Holmberg L, Zimmerman TS. Aberrant multimeric structure of von Willebrand factor in a new variant of von Willebrand’s disease (type IIC). J Clin Invest 1982; 70: 1124-1127
• 7 Mannucci PM, Lombardi R, Pareti FI, Solinas S, Mazzucconi MG, Mariani G. A variant of von Willebrand’s disease characterized by recessive inheritance and missing triplet structure of von Willebrand factor multimers. Blood 1983; 62: 1000-1005
• 8 Hoyer LW, Rizza CR, Tuddenham EG, Carta CA, Armitage H, Rotblat F. Von Willebrand factor multimer patterns in von Willebrand’s disease. Brit J Haematol 1983; 55: 493-507
• 9 Mazurier C, Parquet-Gernez A, Samor B, Goudemand M, Montreuil J. Etude de 1’antigène lié au facteur VIII (FVIIIR:Ag). Purification par chromatographie d’affinité. C R Acad Sci Paris 1979; 288: 1431-1434
• 10 Nilsson IM. Von Willebrand’s disease from 1926-1983. Scand J Haematol 1984; (Suppl. 33) supp 40: 21-43
• 11 Hill FG H, Enayat MS, George AJ. Investigation including VIIIR:Ag multimeric analysis of a large kindred with type IIA von Willebrand’s disease showing a dominant inheritance and similar gene expression in four generations. Thromb Haemostas 1983; 50: 735-739
• 12 Zimmerman TS, Dent JA, Federici ZM, Nilsson IM, Holmberg L, Abildgaard CA, Nannini LH. High resolution Na-DodS04 agarose electrophoresis identifies new molecular abnormalities in von Willebrand’s disease. Clin Res 1984; 32: A501