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Thromb Haemost 1986; 55(02): 263-267
DOI: 10.1055/s-0038-1661533
Original Article
Schattauer GmbH Stuttgart

Visualization of von Willebrand Factor Multimers by Immunoenzymatic Stain Using Avidin-Biotin Peroxidase Complex

M Aihara
The First Department of Internal Medicine, Hirosaki University School of Medicine, Hirosaki, Japan
,
Y Sawada
The First Department of Internal Medicine, Hirosaki University School of Medicine, Hirosaki, Japan
,
K Ueno
The First Department of Internal Medicine, Hirosaki University School of Medicine, Hirosaki, Japan
,
S Morimoto
The First Department of Internal Medicine, Hirosaki University School of Medicine, Hirosaki, Japan
,
Y Yoshida
The First Department of Internal Medicine, Hirosaki University School of Medicine, Hirosaki, Japan
,
M de Serres
1   The Department of Pathology and Center for Thrombosis and Hemostasis, The University of North Carolina, Chapel Hill, NC, USA
,
H A Copper
1   The Department of Pathology and Center for Thrombosis and Hemostasis, The University of North Carolina, Chapel Hill, NC, USA
,
R H Wegner
1   The Department of Pathology and Center for Thrombosis and Hemostasis, The University of North Carolina, Chapel Hill, NC, USA
› Author Affiliations
Further Information

Publication History

Received 07 January 1986

Accepted 18 February 1986

Publication Date:
18 July 2018 (online)

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Summary

A technique for the detection of von Willebrand factor multimers separated by discontinuous SDS agarose electrophoresis has been developed using non-radioactive com-v pounds. The multimeric patterns were visualized by monospecific anti-human vWF:Ag followed by incubation with biotinylated antibody. After addition of avidin-biotin-peroxidase complex, the peroxidase activitiy was detected by 4-chloro-l-naphthol, giving sharp bands with a clear background.

By this method, the differences of vWF : Ag multimers could be easily observed between normal plasma and the plasmas from variant type vWD (IIA, IIB, platelet-type). Large and intermediate multimers were absent in the plasma with vWD type IIA, while only large multimers were absent in the plasma with vWD IIB and platelet-type. The absence of large multimers was also observed in two commercial F VIII preparations having the ratio of vWF/vWF : Ag 0.18 and 0.63. The preparation with the ratio of 0.63 showed the presence of larger intermediate multimers.

Electrophoresis in SDS 1.5% agarose gel revealed triplet structure of each small multimer, and a relative increase of the smallest subband was observed in vWD IIA plasma, platelet-type vWD plasma and commercial F VIII preparations.

The procedures described are easy and safe to perform and are useful for screening or classifying cases with vWD in general laboratories.

Key words

SDS-agarose gel electrophoresis - von Willebrand factor multimers - Immunoenzymatic stain - Avidin-biotin peroxidase complex - von Willebrand’s disease
 

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