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Published on: December 28, 2015
Purification Methods for Recombinant Factor VIII Expressed in Human Liver SK-Hep Cells
Vladimir Granovski1, Mario Soares Abreu-Neto2, Dimas Tadeu Covas3,2
1Center for Cell-based Therapy CTC, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, Brazil. granovski.v@gmail.com.
Abstract:
Coagulation factor VIII is one of the largest proteins attempted to be expressed in recombinant form. A very complex and labile protein which has a very short half-live and need a fast and efficient purification chain. Here, we describe a simple purification sequence using multimodal Capto MMC, affinity FVIII select and ion exchange SP-Fastflow chromatography steps without subjecting the target molecule to mechanical and temperature stress, separating impurities from rFVIII using net charge, hydrophobicity, and affinity of the molecules.
