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Purification of influenza virus‐like particles using sulfated cellulose membrane adsorbers
Author(s) -
B Carvalho Sofia,
Fortuna A Raquel,
Wolff Michael W,
Peixoto Cristina,
M Alves Paula,
Reichl Udo,
JT Carrondo Manuel
Publication year - 2018
Publication title -
journal of chemical technology and biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 117
eISSN - 1097-4660
pISSN - 0268-2575
DOI - 10.1002/jctb.5474
Subject(s) - ultrafiltration (renal) , membrane , chromatography , downstream processing , yield (engineering) , chemistry , ion exchange , immunogenicity , elution , fraction (chemistry) , nanofiltration , antigen , biochemistry , materials science , biology , organic chemistry , ion , metallurgy , genetics
ABSTRACT BACKGROUND Vaccines based on virus‐like particles (VLPs) are an alternative to inactivated viral vaccines that combine good safety profiles with strong immunogenicity. In order to be economically competitive, efficient manufacturing is required, in particular downstream processing, which often accounts for major production costs. This study describes the optimization and establishment of a chromatography capturing technique using sulfated cellulose membrane adsorbers (SCMA) for purification of influenza VLPs. RESULTS Using a design of experiments approach, the critical factors for SCMA performance were described and optimized. For optimal conditions (membrane ligand density: 15.4 µmol cm −2 , salt concentration of the loading buffer: 24 mmol L ‐1 NaCl, and elution buffer: 920 mmol L ‐1 NaCl, as well as the corresponding flow rates: 0.24 and 1.4 mL min −1 ), a yield of 80% in the product fraction was obtained. No loss of VLPs was detected in the flowthrough fraction. Removal of total protein and DNA impurities were higher than 89% and 80%, respectively. CONCLUSION Use of SCMA represents a significant improvement compared with conventional ion exchanger membrane adsorbers. As the method proposed is easily scalable and reduces the number of steps required compared with conventional purification methods, SCMA could qualify as a generic platform for purification of VLP‐based influenza vaccines. © 2017 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.