Optimization of von Willebrand factor multimer analysis in vertical mini-gel electrophoresis systems: A rapid procedure

• Published in: Thrombosis research 2019-03, Vol.175, p.76-83
• Main Authors: Thomazini, Camila Martos, Soares, Rosangela de Paula Silva, da Rocha, Tania Rubia Flores, Sachetto, Ana Teresa Azevedo, Santoro, Marcelo Larami
• Format: Article
• Language: English
• Subjects: ADAMTS13; Bleeding; Diagnosis; Electrophoresis; Proteolysis
• ISSN: 0049-3848; 1879-2472
• DOI: 10.1016/j.thromres.2019.01.018

Von Willebrand disease (VWD) is a common cause of bleeding worldwide. Analysis of von Willebrand factor (VWF) multimer distribution (VWF:MD) is essential to properly classify and treat different types of VWD, and it is performed using a SDS agarose gel electrophoresis followed by Western blotting, a handmade technique that demands days to be completed and requires skillful execution. Aiming both to facilitate gel production and to shorten the preparation time, we developed an uncomplicated technique to provide agility in the analysis of VWF:MD, so that it can be easily accomplished in the routine practice of hemostasis laboratories. On that account, we used a commercial vertical mini-gel electrophoresis system for SDS-PAGE and a semi-dry transfer system, which allowed us to analyze VWF:MD of various samples in a period shorter than 12 h. This technique differentiated VWF:MD in human and animal plasmas under normal, congenital and acquired (experimental envenomation by Bothrops jararaca snake) conditions. This optimized method is cheap, rapid, reproducible, easy to be performed, and uses electrophoresis and Western blotting systems available in most laboratories. All these advantages encourage hemostasis professionals to use it in their routine practices. In order to facilitate the setup and accomplishment of the whole procedure step by step, videos were appended to the article. •Analysis of von Willebrand factor multimer distribution was optimized to be performed in 12 h.•A commercial vertical mini-gel electrophoresis system and a semi-dry transfer unit were employed.•Differentiation of normal and pathological multimer distribution was achieved in human and animal samples.•This simplified procedure can be set up in research and clinical hemostasis laboratories.