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Influence of cytoskeleton organization on recombinant protein expression by CHO cells.
Pourcel, Lucille; Buron, Flavien; Arib, Ghislaine; Le Fourn, Valérie; Regamey, Alexandre; Bodenmann, Iris; Girod, Pierre-Alain; Mermod, Nicolas.
Afiliación
  • Pourcel L; Department of Fundamental Microbiology, Institute of Biotechnology, University of Lausanne, Lausanne, Switzerland.
  • Buron F; Department of Fundamental Microbiology, Institute of Biotechnology, University of Lausanne, Lausanne, Switzerland.
  • Arib G; Selexis SA, Geneva, Switzerland.
  • Le Fourn V; Selexis SA, Geneva, Switzerland.
  • Regamey A; Selexis SA, Geneva, Switzerland.
  • Bodenmann I; Selexis SA, Geneva, Switzerland.
  • Girod PA; Selexis SA, Geneva, Switzerland.
  • Mermod N; Department of Fundamental Microbiology, Institute of Biotechnology, University of Lausanne, Lausanne, Switzerland.
Biotechnol Bioeng ; 117(4): 1117-1126, 2020 04.
Article en En | MEDLINE | ID: mdl-31956990
In this study, we assessed the importance of cytoskeleton organization in the mammalian cells used to produce therapeutic proteins. Two cytoskeletal genes, Actin alpha cardiac muscle 1 (ACTC1) and a guanosine triphosphate GTPase-activating protein (TAGAP), were found to be upregulated in highly productive therapeutic protein-expressing Chinese hamster ovary (CHO) cells selected by the deprivation of vitamin B5. We report here that the overexpression of the ACTC1 protein was able to improve significantly recombinant therapeutic production, as well as to decrease the levels of toxic lactate metabolic by-products. ACTC1 overexpression was accompanied by altered as well as decreased polymerized actin, which was associated with high protein production by CHO cell cultured in suspension. We suggest that the depolymerization of actin and the possible modulation of integrin signaling, as well as changes in basal metabolism, may be driving the increase of protein secretion by CHO cells.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Citoesqueleto de Actina / Proteínas Recombinantes / Actinas 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: Citoesqueleto de Actina / Proteínas Recombinantes / Actinas Límite: Animals Idioma: En Revista: Biotechnol Bioeng Año: 2020 Tipo del documento: Article País de afiliación: Suiza Pais de publicación: Estados Unidos