Using the “OMICS” Technologies as Complementary Tools to Study the Molecular Mechanisms Involved with the Adaptation of Myeloma Cell Line to Protein-Free Medium
Production of recombinant therapeutic proteins, especially monoclonal antibodies (Mab), in myeloma cell lines represents a significant segment of the pharmaceutical market, and therefore striving for increased productivity of these lines represents a major investment of resources. The elucidation of biologically important markers for the adaptation of NS0 myeloma cell line to protein-free medium and the recombinant protein production are a major emphasis of our research. These markers could potentially be used in a variety of ways to improve culture conditions, including active approaches to agonize/antagonize important pathways within a medium formulation or diagnostic approaches indicative of improved conditions during the culture. In this work, we used two-dimensional electrophoresis/mass spectrometry and the iTRAQ technology to analyze different protein levels in adapted and non-adapted NS0 myeloma cell line. Several proteins with differential expression profile were characterized and quantified. Changes in lactate production rate with respect to glucose consumption rate were observed according to the changes observed by proteomic. Carbohydrate metabolism, protein synthesis and membrane transport were the principal pathways that change after the adaptation by proteomic analysis. The same results were obtained using flux balance analysis in a murine metabolic network with selected medium conditions.
KeywordsMetabolic Network Flux Balance Analysis Recombinant Therapeutic Protein Lactate Production Rate Intracellular Metabolite Concentration
- Castillo AJ, Victores S, Rojas L, Faife E, Rabasa Y, Pence BL, et al. Adaptation and selection of NS0 myeloma cell lines producing recombinant monoclonal antibodies in protein-free medium. In: Lindner-Osson E, et al, editor. Animal cell technology: from target to market. Kluwer, Dordrecht; 2001. pp. 160–3.CrossRefGoogle Scholar
- de la Luz-Hernández KR, Rojas-del Calvo L, Rabasa-Legón Y, Lage-Castellanos A, Castillo-Vitlloch A, Díaz, J, Gaskell S. Metabolic and proteomic study of NS0 myeloma cell line following the adaptation to protein-free medium. Journal of Proteomics 2008;71:133–147.Google Scholar
- de la Luz-Hernández KR, Rojas-del Calvo L, Victores-Sarasola S, Lage-Castellanos A, Eyers C, Hart H, et al. Proteomic analysis of the adaptation of the host NS0 myeloma cell line to a protein-free medium. Biotecnologia Aplicada, 2007;24:215–223.Google Scholar
- Roque L, Mateo C, Lombardero J, Mustelier A, Fernández A, Sosa K, et al. Humanization of predicted T-cell epitopes reduces the immunogenecity of chimeric antibodies: new evidences supporting a simple method. Hybridoma 2003;22:245–57.Google Scholar