Luminographic Detection of von Willebrand Factor Multimers in Agarose Gels and on Nitrocellulose Membranes

U Budde; R Schneppenheim; H Plendl; J Dent; Z M Ruggeri; T S Zimmerman

doi:10.1055/s-0038-1645215

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1990; 63(02): 312-315
DOI: 10.1055/s-0038-1645215

Original Article

Schattauer GmbH Stuttgart

Luminographic Detection of von Willebrand Factor Multimers in Agarose Gels and on Nitrocellulose Membranes

U Budde

¹The Blood Transfusion Service AK Harburg, Hamburg, FRG

,

R Schneppenheim

²The Dept. of Pediatrics, University Clinic, Kiel, FRG

,

H Plendl

³The Dept. of Human Genetics, University Clinic, Kiel, FRG

,

J Dent

⁴The Scripps Clinic and Research Foundation, La Jolla, CA, USA

,

Z M Ruggeri

⁴The Scripps Clinic and Research Foundation, La Jolla, CA, USA

,

T S Zimmerman

⁴The Scripps Clinic and Research Foundation, La Jolla, CA, USA

› Author Affiliations

Abstract

PDF Download

Summary

Two methods for visualization of vWf multimers were compared with respect to sensitivity and detection of normal vWf and vWd variants IIA, IIB, IIC, IID, HE, and HE Autoradiography and luminography after electrotransfer of vWf multimers onto nitrocellulose showed comparable sensitivity with vWf: Ag detectable after 1:500 dilution of normal plasma. The least sensitive method was luminography in agarose gels with vWf: Ag detectable after 1:300dilution of normal plasma. No difference existed in the banding patterns of plasmas from patients with variant vWd.

PDF (366 kb)

References

References
1 Ruggeri ZM, Zimmerman TS. Von Willebrand factor and von Willebrand disease. Blood 1987; 70: 895-904
2 Ruggeri ZM, Zimmerman TS. The complex multimeric composition of factor VIII von Willebrand factor. Blood 1981; 57: 1140-1143
3 Brosstad F, Kjoenniksen I, Roenning B, Stormorken H. Visualization of von Willebrand factor multimers by enzyme-conjugated second antibodies. Thromb Haemostas 1986; 55: 276-278
4 Bukh A, Ingerslev J, Stenbjerg S, Hundahl Moeller NR. The multimeric structure of plasma F VIII: RAg studied by electroelution and immunoperoxidase detection. Thromb Res 1984; 43: 579-584
5 Lombardi R, Gelfi C, Righetti PG, Lattuada A, Mannucci PM. Electroblot and immunoperoxidase staining for rapid screening of the abnormalities of the multimeric structure of von Willebrand factor in von Willebrand’s disease. Thromb Haemostas 1986; 55: 246-249
6 Aihara M, Sawada Y, Ueno K, Morimoto S, Yoshida Y, De Serres M, Cooper HA, Wagner RH. Visualization of von Willebrand factor multimers by immunoenzymatic stain using avidin-biotin peroxidase complex. Thromb Haemostas 1986; 55: 263-267
7 Schneppenheim R, Plendl H, Budde U. Luminography – an alternative assay for detection of von Willebrand factor multimers. Thromb Haemostas 1988; 60: 133-136
8 Mannucci PM, Lombardi R, Castaman G, Dent JA, Lattuada A, Rodeghiero F, Zimmerman TS. Von Willebrand disease Vicenza with larger-than-normal (supranormal) von Willebrand factor multimers. Blood 1988; 71: 65-70
9 Moake JL, Rudy CK, Troll JH, Weinstein MJ, Collannino NM, Azocar J, Seder RH, Hong SL, Deykin D. Usually large plasma factor multimers in chronic relapsing thrombotic thrombocytopenic purpura. N Engl J Med 1982; 307: 1432-1435
10 Budde U, Schafer G, Muller N, Egli H, Dent J, Ruggeri ZM, Zimmerman TS. Acquired von Willebrand’s disease in the myeloproliferative syndrome. Blood 1984; 64: 981-985
11 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
12 Enayat MS, Hill FG H. Analysis of the complexity of the multimeric structure of the factor VIII related antigen/von Willebrand protein using a modified electrophoretic technique. J Clin Pathol 1983; 36: 915-919