von Willebrand Factor Multimer Analysis Using a Sensitive Peroxidase Staining Method

Summary

A peroxidase staining method for von Willebrand factor (vWF) multimer analysis was modified for comparison with an autoradiographic method. This method consists of SDS-agarose electrophoresis followed by blotting to a nitrocellulose membrane and a sensitive peroxidase staining method. The resolution of vWF multimers on the nitrocellulose membrane is comparable to that on the conventional autoradiography. Results can be obtained in 3 days. This nonradioisotopic method will be useful for the determination of the type of von Willebrand’s disease in clinical laboratories.

Keywords

von Willebrand factor multimer analysis

• References

• 1 Ruggeri ZM, Zimmerman TS. von Willebrand factor and von Willebrand disease. Blood 1987; 70: 895-904
• 2 Abildgaard CF. Progress and problems in hemophilia and von Willebrand’s disease. In: Advances in Pediatrics. (; 31). Bamess LA. (ed) Year Book Medical Publishers; Chicago: 1984: 137-177
• 3 Ruggeri ZM, Mannucci PM, Lombardi R, Federici AB, Zimmerman TS. Multimeric composition of factor VUI/von Willebrand factor following administration of DDAVP: Implications for pathophysiology and therapy of von Willebrand’s disease subtypes. Blood 1982; 59: 1272-1278
• 4 Holmberg L, Nilsson IM, Borge L, Gunnarsson M, Sjorin E. Platelet aggregation induced by l-desamino-8-D-arginine vasopressin (DDAVP) in type IIB von Willebrand’s disease. N Engl J Med 1983; 309: 816-821
• 5 Abildgaard CF, Suzuki Z, Harrison J, Jefcoat K, Zimmerman TS. Serial studies in von Willebrand’s disease: Variability versus “variants”. Blood 1980; 56: 712-716
• 6 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 VUI-related antigen (type IID). Blood 1984; 63: 1369-1371
• 7 Zimmerman TZ, Robert JR, Ruggeri ZM. Factor VUI-related antigen: Characterization by electrophoretic technique. In: The Hemophilias, Methods in Hematology. (; 5). Bloom AL. (ed) Churchill Livingstone; Edinburgh: 1982: 81-91
• 8 Towbin H, Staehelin T, Gordon J. Electrophoretic transfer or proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 1979; 76: 4350-4354
• 9 Johnson DA, Gautsch JW, Sportsman JR, Elder JH. Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal Techn 1984; 1: 3-8
• 10 Furlong BL, Peake IR. An electroblotting technique for the detection of factor VUI/von Willebrand factor multimers in plasma. Br J Haematol. 1983: 641-653
• 11 Zaleski A, Henriksen RA. Visualization of the multimeric structure of von Willebrand factor using a peroxidase-conjugated second antibody. J Lab Clin Med 1986; 107: 172-175
• 12 Bukh A, Ingerslev J, Stenbjerg S, Moller NP H. The multimeric structure of plasma F VIII: RAg studied by electroelution and immunoperoxidase detection. Thromb Res 1986; 43: 579-584
• 13 Takahashi H, Tatewaki W, Hanano M, Shibata A. Platelet-type von Willebrand disease platelet aggregating factor: A novel functional assay of von Willebrand factor. Thromb Res 1987; 48: 475-485
• 14 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