Are the multimers of von Willebrand factor important for the confirmation of the diagnosis of certain vWD subtypes? What is your experience with their assessment? Can you recommend your farourite method?
Hi Dr Peter,
Yes, von Willebrand factor (VWF) multimer pattern is very important for diagnosis and classification of different type of VWD. Type 2 VWD can further be classified into its subtypes on the basis of VWF multimer pattern. The best method I know to detect the full range of VWF multimers is horizontal SDS-agarose gel electrophoresis (0.8% stacking and 1.6% running gel) using high gelling temperature (SeaKam) agarose and cool plated electrophoresis chamber (Pharmacia Multiphor II) followed by vertical blotting. I would recommend for primary Ab from DAKO (Rabbit anti-human) and secondary Ab from BioRad (Goat anti-Rabbit-HRP). Dr Budde’s lab in Hamburg is one of the best laboratories in the world for VWF multimeric analysis. For more detail you may have look on our recently published articles (Thromb Haemost. 2013 Jan;109(1):39-46 and Thromb Haemost. 2013 Apr 8;109(4):652-60)
Hi Dr. Fridos Ahmad,
Thank you very much for your comment to my query. I would like to know your opinion regarding two other related questions:
1 Is luminescence method currently the most sensitive method for the analysis of vWF multimers?
2 Is there software to determine the molecular weight of vWF multimers evaluated by densitometry?
Hi Peter,
Yes, it is sensitive. But, luminescence method may help in case of very low level of VWF, and in normals it may be difficult to read the multimers as the staining will give dense, merged bands.
In our experience, immunoenzymztic method is the best. This will allow you to read HMW multimers clearly.
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