Purification of influenza virus-like particles using sulfated cellulose membrane adsorbers

Journal of Chemical Technology and Biotechnology
Sofia B CarvalhoManuel Jt Carrondo

Abstract

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. 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. 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 w...Continue Reading

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Citations

Jan 1, 2019·Biotechnology and Bioengineering·Hayley K Charlton HumeLinda H L Lua
Nov 28, 2019·Expert Opinion on Biological Therapy·Mafalda G MoleirinhoCristina Peixoto
May 21, 2019·Biotechnology Journal·Sofia B CarvalhoCristina Peixoto
Sep 10, 2020·Asian Journal of Pharmaceutical Sciences·Lanxiang HuangFubing Wang
Apr 30, 2021·Biotechnology and Bioengineering·Sofia B CarvalhoRicardo J S Silva
Jul 3, 2021·Membranes·Abayomi Babatunde AlayandeEuntae Yang
Jul 6, 2021·Biotechnology and Bioengineering·Thomas BissingerUdo Reichl

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Methods Mentioned

BETA
dynamic light scattering
light scattering
Protein Assay

Software Mentioned

MODDE Pro
MODDE Optimizer
ExPASy ProtParam

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