Enhancing the purification of Lentiviral vectors for clinical applications

A. S. Moreira, T. Q. Faria, Joana G Oliveira, A. Kavara, M. Schofield, T. Sanderson, M. Collins, R. Gantier, P. M. Alves, M. J.T. Carrondo, C. Peixoto

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

Lentiviral vectors have been increasingly used as a tool for gene and cell therapies since they can stably integrate the genome in dividing and nondividing cells. The development of efficient, fast, and robust downstream processes that cope with the low stability of this virus is slowing down the clinical-to-market transition. A purification process comprised by four-unit operation was developed and improved. Several clarification filters with different materials were evaluated for virus recovery and throughput. Anion exchange chromatography was optimized by tuning the critical operation conditions for Mustang™ Q and Sartobind™ Q membrane chromatography devices using Design of Experiments methodology. Mustang Q demonstrated better performance with recovery yields close to 90%. Good scalability and robustness were found running Mustang Q at different scales up to 5 mL capsule. Flat sheet cassettes and hollow fiber with different molecular weight cut-offs were evaluated for concentration and formulation and recovery yields above 60% were obtained for both supports operated under different experimental conditions. The sterile filtration step was successfully performed and allowing a global virus recovery up to 45%. This work unravels strategies to enhance lentivirus vector purification process which may accelerate the development of therapeutic products based on lentiviral vectors.

Original languageEnglish
Article number118598
JournalSeparation and Purification Technology
Volume274
DOIs
Publication statusPublished - 1 Nov 2021

Keywords

  • Chromatography
  • Design of experiments
  • Downstream processing
  • Lentiviral vector
  • Tangential flow filtration

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