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

Sofia B. Carvalho, A. Raquel Fortuna, Michael W. Wolff, Cristina Peixoto, Paula M. Alves, Udo Reichl, Manuel J.T. Carrondo

Research output: Contribution to journalArticle

10 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1988-1996
Number of pages9
JournalJournal of Chemical Technology and Biotechnology
Volume93
Issue number7
DOIs
Publication statusPublished - 1 Jul 2018

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virus particle
influenza
Orthomyxoviridae
Viruses
Cellulose
Virion
Purification
purification
cellulose
membrane
Membranes
Vaccines
vaccine
Buffers
Viral Vaccines
Inactivated Vaccines
Ion exchangers
Influenza Vaccines
production cost
Chromatography

Keywords

  • downstream processing
  • membrane adsorption chromatography
  • sulfated cellulose
  • vaccine production
  • virus-like particles

Cite this

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title = "Purification of influenza virus-like particles using sulfated cellulose membrane adsorbers",
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.",
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Purification of influenza virus-like particles using sulfated cellulose membrane adsorbers. / Carvalho, Sofia B.; Fortuna, A. Raquel; Wolff, Michael W.; Peixoto, Cristina; Alves, Paula M.; Reichl, Udo; Carrondo, Manuel J.T.

In: Journal of Chemical Technology and Biotechnology, Vol. 93, No. 7, 01.07.2018, p. 1988-1996.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Purification of influenza virus-like particles using sulfated cellulose membrane adsorbers

AU - Carvalho, Sofia B.

AU - Fortuna, A. Raquel

AU - Wolff, Michael W.

AU - Peixoto, Cristina

AU - Alves, Paula M.

AU - Reichl, Udo

AU - Carrondo, Manuel J.T.

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N2 - 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.

AB - 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.

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