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Overexpression of transcription factor Foxa1 and target genes remediate therapeutic protein production bottlenecks in Chinese hamster ovary cells.
Berger, Audrey; Le Fourn, Valérie; Masternak, Jacqueline; Regamey, Alexandre; Bodenmann, Iris; Girod, Pierre-Alain; Mermod, Nicolas.
Afiliación
  • Berger A; Department of Fundamental Microbiology, Institute of Biotechnology, University of Lausanne, Lausanne, Switzerland.
  • Le Fourn V; Present address: Laboratory of Microsystems LMIS4, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.
  • Masternak J; Selexis SA, Geneva, Switzerland.
  • Regamey A; Department of Fundamental Microbiology, Institute of Biotechnology, University of Lausanne, Lausanne, Switzerland.
  • Bodenmann I; Selexis SA, Geneva, Switzerland.
  • Girod PA; Selexis SA, Geneva, Switzerland.
  • Mermod N; Selexis SA, Geneva, Switzerland.
Biotechnol Bioeng ; 117(4): 1101-1116, 2020 04.
Article en En | MEDLINE | ID: mdl-31956982
Despite extensive research conducted to increase protein production from Chinese hamster ovary (CHO) cells, cellular bottlenecks often remain, hindering high yields. In this study, a transcriptomic analysis led to the identification of 32 genes that are consistently upregulated in high producer clones and thus might mediate high productivity. Candidate genes were associated with functions such as signaling, protein folding, cytoskeleton organization, and cell survival. We focused on two engineering targets, Erp27, which binds unfolded proteins and the Erp57 disulfide isomerase in the endoplasmic reticulum, and Foxa1, a pioneering transcription factor involved in organ development. Erp27 moderate overexpression increased production of an easy-to-express antibody, whereas Erp27 and Erp57 co-overexpression increased cell density, viability, and the yield of difficult-to-express proteins. Foxa1 overexpression increased cell density, cell viability, and easy- and difficult-to-express protein yields, whereas it decreased reactive oxygen species late in fed-batch cultures. Foxa1 overexpression upregulated two other candidate genes that increased the production of difficult- and/or easy-to-express proteins, namely Ca3, involved in protecting cells from oxidative stress, and Tagap, involved in signaling and cytoskeleton remodeling. Overall, several genes allowing to overcome CHO cell bottlenecks were identified, including Foxa1, which mediated multiple favorable metabolic changes that improve therapeutic protein yields.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Recombinantes / Factor Nuclear 3-alfa del Hepatocito / Ingeniería Celular Tipo de estudio: Prognostic_studies Límite: Animals Idioma: En Revista: Biotechnol Bioeng Año: 2020 Tipo del documento: Article País de afiliación: Suiza Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Recombinantes / Factor Nuclear 3-alfa del Hepatocito / Ingeniería Celular Tipo de estudio: Prognostic_studies Límite: Animals Idioma: En Revista: Biotechnol Bioeng Año: 2020 Tipo del documento: Article País de afiliación: Suiza Pais de publicación: Estados Unidos