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A new T-antigen negative HEK293 cell line with improved AAV productivity
  • +11
  • Paco Pino,
  • Coralie Croissant,
  • Joshua Armitano,
  • Bertrand Lazuech,
  • Danijel Švec,
  • Cyril Pugin,
  • Anaïs Guesdon,
  • Louise Bryan,
  • Antonio Castro,
  • Léa Neuhaus,
  • Giulia Fonti,
  • Jacopo Martinis,
  • Maria Wurm,
  • Florian M. Wurm
Paco Pino
ExcellGene SA

Corresponding Author:paco.pino@excellgene.com

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Coralie Croissant
ExcellGene SA
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Joshua Armitano
ExcellGene SA
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Bertrand Lazuech
ExcellGene SA
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Danijel Švec
ExcellGene SA
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Cyril Pugin
ExcellGene SA
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Anaïs Guesdon
ExcellGene SA
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Louise Bryan
ExcellGene SA
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Antonio Castro
ExcellGene SA
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Léa Neuhaus
ExcellGene SA
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Giulia Fonti
ExcellGene SA
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Jacopo Martinis
ExcellGene SA
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Maria Wurm
ExcellGene SA
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Florian M. Wurm
ExcellGene SA
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Abstract

Viral vectors for gene therapy, such as recombinant Adeno-Associated Viruses (rAAV), are produced in Human Embryonic Kidney (HEK) 293 cells. However, the presence of the SV40 T-antigen-encoding CDS SV40GP6 and SV40GP7 in the HEK293T genome raises safety issues when these cells are used in manufacturing for clinical purposes. We developed a new T-antigen-negative HEK cell line from ExcellGene’s proprietary HEKExpress®, using the CRISPR-Cas9 strategy. We obtained a high number of clonally-derived cell populations and all of them were demonstrated T-antigen negative. Stability study and AAV production evaluation showed that the deletion of the T-antigen-encoding locus did not impact neither cell growth nor viability nor productivity. The resulting CMC-compliant cell line, named HEKzeroT®, is able to produce high AAV titers, from small to large scale.
17 Jan 2023Submitted to Biotechnology and Bioengineering
20 Jan 2023Assigned to Editor
20 Jan 2023Submission Checks Completed
20 Jan 2023Review(s) Completed, Editorial Evaluation Pending
22 Jan 2023Reviewer(s) Assigned