Towards the molecular characterization of the stable producer phenotype of recombinant antibody-producing NS0 myeloma cells

Y. Prieto, L. Rojas, L. Hinojosa, I. González, D. Aguiar, K. De La Luz, A. Castillo, R. Pérez

Producción científica: Contribución a una revistaArtículorevisión exhaustiva

9 Citas (Scopus)

Resumen

The loss of heterologous protein expression is one of the major problems faced by industrial cell line developers and has been reported by several authors. Therefore, the understanding of the mechanisms involved in the generation of stable and high producer cell lines is a critical issue, especially for those processes based on long term continuous cultures. We characterized two recombinant NS0 myeloma cell lines expressing Nimotuzumab, a humanized anti-human epidermal growth factor receptor (EGFR) antibody. The hR3/H7 clone is a stable producer obtained from the unstable hR3/t16 clone. The unstable clone was characterized by a bimodal distribution of intracellular immunoglobulin staining using flow cytometry. Loss of antibody production was due to the emergence of a non-producer cell subpopulation that increased with cell generation number. Immunoglobulin heavy chain (HC) and light chain (LC) ratio (HC/LC) was lower for the unstable phenotype. Proteomic maps using two dimensional gel electrophoresis (2DE) were obtained for both clones, at initial cell culture time and after 40 generations. Fifteen proteins potentially associated with the phenomenon of production stability were identified. The hR3/H7 stable clone showed an up-regulated expression pattern for most of these proteins. The regulation of recombinant antibody production by the host NS0 myeloma cell line most likely involves simultaneously cellular processes such as DNA transcription, mRNA processing, protein synthesis and folding, vesicular transport, glycolysis and energy production, according to the proteins identified in the present proteomic study.

Idioma originalInglés
Páginas (desde-hasta)351-362
Número de páginas12
PublicaciónCytotechnology
Volumen63
N.º4
DOI
EstadoPublicada - ago. 2011
Publicado de forma externa

Huella

Profundice en los temas de investigación de 'Towards the molecular characterization of the stable producer phenotype of recombinant antibody-producing NS0 myeloma cells'. En conjunto forman una huella única.

Citar esto