tetano
Editor, Senior Moderator
J Chem Technol Biotechnol. 2018 Jul;93(7):1988-1996. doi: 10.1002/jctb.5474. Epub 2017 Dec 16.
[h=1]Purification of influenza virus-like particles using sulfated cellulose membrane adsorbers.[/h] B Carvalho S[SUP]1,[/SUP][SUP]2[/SUP], Fortuna AR[SUP]3[/SUP], Wolff MW[SUP]3,[/SUP][SUP]4[/SUP], Peixoto C[SUP]1,[/SUP][SUP]2[/SUP], M Alves P[SUP]1,[/SUP][SUP]2[/SUP], Reichl U[SUP]3,[/SUP][SUP]5[/SUP], Jt Carrondo M[SUP]1,[/SUP][SUP]6[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] [h=4]BACKGROUND:[/h] 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.
[h=4]RESULTS:[/h] Using a design of experiments approach, the critical factors for SCMA performance were described and optimized. For optimal conditions (membrane ligand density: 15.4 ?mol cm[SUP]-2[/SUP], salt concentration of the loading buffer: 24 mmol L[SUP]-1[/SUP] NaCl, and elution buffer: 920 mmol L[SUP]-1[/SUP] NaCl, as well as the corresponding flow rates: 0.24 and 1.4 mL min[SUP]-1[/SUP]), 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.
[h=4]CONCLUSION:[/h] 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.
[h=4]KEYWORDS:[/h] downstream processing; membrane adsorption chromatography; sulfated cellulose; vaccine production; virus‐like particles
PMID: 30008506 PMCID: PMC6033026 DOI: 10.1002/jctb.5474
[h=1]Purification of influenza virus-like particles using sulfated cellulose membrane adsorbers.[/h] B Carvalho S[SUP]1,[/SUP][SUP]2[/SUP], Fortuna AR[SUP]3[/SUP], Wolff MW[SUP]3,[/SUP][SUP]4[/SUP], Peixoto C[SUP]1,[/SUP][SUP]2[/SUP], M Alves P[SUP]1,[/SUP][SUP]2[/SUP], Reichl U[SUP]3,[/SUP][SUP]5[/SUP], Jt Carrondo M[SUP]1,[/SUP][SUP]6[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] [h=4]BACKGROUND:[/h] 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.
[h=4]RESULTS:[/h] Using a design of experiments approach, the critical factors for SCMA performance were described and optimized. For optimal conditions (membrane ligand density: 15.4 ?mol cm[SUP]-2[/SUP], salt concentration of the loading buffer: 24 mmol L[SUP]-1[/SUP] NaCl, and elution buffer: 920 mmol L[SUP]-1[/SUP] NaCl, as well as the corresponding flow rates: 0.24 and 1.4 mL min[SUP]-1[/SUP]), 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.
[h=4]CONCLUSION:[/h] 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.
[h=4]KEYWORDS:[/h] downstream processing; membrane adsorption chromatography; sulfated cellulose; vaccine production; virus‐like particles
PMID: 30008506 PMCID: PMC6033026 DOI: 10.1002/jctb.5474