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von W illebrand F actor multimers profiling with a semi‐automated system
Author(s) -
Pasotti Fabio,
Martini Giuliana,
Caimi Luigi,
Ricotta Doris
Publication year - 2013
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201200538
Subject(s) - von willebrand factor , von willebrand disease , blot , genotyping , computational biology , computer science , microbiology and biotechnology , chemistry , biology , immunology , genetics , gene , genotype , platelet
Abnormalities in plasma von Willebrand factor (v WF ) concentration and function result in von Willebrand disease (v WD ). The diagnosis requires a battery of tests such as screening procedures, confirmatory tests, phenotypic characterization, and genotyping. The phenotypic testing (multimer pattern analysis) is important in order to subclassify the hereditary and the acquired forms of v WD . Only few laboratories are skilled to perform this analysis. The extreme range of protein size from 250 kDa monomer to over 20 000 kDa multimers requires a time‐consuming procedure (3–4 days) and presents many technical difficulties. To standardize the method and to overcome technical difficulties, we developed a rapid and sensitive semi‐automated method to visualize the multimeric structure of v WF . The semi‐automated method we present performs the electrophoresis of patient's plasma in 120 min on a precast gel. Gels are suitable for the G 26 Interlab instrumentation. After gel blotting, the method allows visualization of the v WF multimer pattern directly on the membrane. We reduced the time required from 72 to 8 h and we propose this test for the first level screening of v WF multimer deficiency.