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Demonstration of a robust high cell density transient CHO platform yielding mAb titers of up to 2 g/L without medium exchange.
Wu, Rigumula; Kahl, Danielle M; Kloberdanz, Ronald; Rohilla, Kushal J; Balasubramanian, Sowmya.
Afiliación
  • Wu R; Department of Cell Culture and Bioprocess Operations, Genentech, Inc, San Francisco, California, USA.
  • Kahl DM; Department of Cell Culture and Bioprocess Operations, Genentech, Inc, San Francisco, California, USA.
  • Kloberdanz R; Department of Cell Culture and Bioprocess Operations, Genentech, Inc, San Francisco, California, USA.
  • Rohilla KJ; Department of Cell Culture and Bioprocess Operations, Genentech, Inc, San Francisco, California, USA.
  • Balasubramanian S; Department of Cell Culture and Bioprocess Operations, Genentech, Inc, San Francisco, California, USA.
Biotechnol Prog ; 40(3): e3435, 2024.
Article en En | MEDLINE | ID: mdl-38329375
ABSTRACT
Biopharmaceuticals like therapeutic monoclonal antibodies (mAbs) and other derived proteins are popular for treating various diseases. Transient gene expression (TGE) is typically used as a fast yet efficient method to generate moderate amounts of material. It has been used to support early stage research and discovery processes. Introduction of a robust high yielding and predictive TGE platform in Chinese hamster ovary (CHO) is crucial. It maintains the consistency in cell lines and processes throughout the early drug discovery and downstream manufacturing processes. This helps researchers to identify the issues at an early stage for timely resolution. In this study, we have demonstrated a simple high-titer platform for TGE in CHO based on a dilution process of seeding cells. We achieved titers ranging from 0.8 to 1.9 g/L for eight model mAbs at three scales (1, 30, 100 mL) in 10 days using our new platform. The ability to seed by dilution significantly streamlined the process and dramatically enhanced platform throughput. We observed a modest reduction in titer ranging from 11% to 28% when cells were seeded using dilution compared to when cells were seeded using medium exchange. Further studies revealed that carry over of spent medium into transfection negatively affected the DNA uptake and transcription processes, while the translation and secretion was minimally impacted. In summary, our transient CHO platform using cells prepared by dilution at high densities can achieve high titers of up to 1.9 g/L, which can be further improved by targeting the bottlenecks of transfection and transcription.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Cricetulus / Anticuerpos Monoclonales Tipo de estudio: Prognostic_studies Límite: Animals Idioma: En Revista: Biotechnol Prog Asunto de la revista: BIOTECNOLOGIA Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Cricetulus / Anticuerpos Monoclonales Tipo de estudio: Prognostic_studies Límite: Animals Idioma: En Revista: Biotechnol Prog Asunto de la revista: BIOTECNOLOGIA Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos