ANALYSIS OF VONWILLEBRAND-FACTOR MULTIMERS USING A COMMERCIALLY AVAILABLE ENHANCED CHEMILUMINESCENCE KIT

Aims-To develop a rapid, sensitive, and safe method for the analysis o f von Willebrand factor (vWf) multimers in plasma or platelet lysates. Method-Analysis of vWf multimers was carried out by sodium dodecyl su lphate-agarose discontinuous gel electrophoresis followed by protein t ransfer to nitrocellulose membranes by western blotting. Blots were pr obed using horseradish peroxidase (HRP) conjugated rabbit anti-vWf; vi sualisation of vWf multimers was achieved using a commercially availab le enhanced chemiLuminescence (ECL) kit for detecting HRP labelled ant ibodies on western blots. Results-Electrophoretic transfer of vWf mult imers to nitrocellulose membranes, including the higher molecular weig ht forms, was achieved satisfactorily and there was good resolution of individual multimer bands and of the triplet subband structure. Type II vWD variants were readily identifiable. The use of ECL conferred a high degree of sensitivity to the method and the end result on autorad iography film provided a permanent record which did not fade and which was suitable for scanning densitometry. Conclusion-The method for vWf multimer analysis described here is sensitive, simple to carry out, u ses minimal amounts of reagents, produces results within 48 hours, and does not require the use of potentially hazardous radioactive materia ls or carcinogenic enzyme substrates.