Using the “OMICS” Technologies as Complementary Tools to Study the Molecular Mechanisms Involved with the Adaptation of Myeloma Cell Line to Protein-Free Medium

  • K.R. de la Luz-Hernández
  • Y. Rabasa-Legón
  • A. Lage-Castellanos
  • A. Castillo-Vitlloch
  • L. Castellanos-Serra
  • J. Díaz-Brito
  • S. Gaskell
Conference paper
Part of the ESACT Proceedings book series (ESACT, volume 5)

Abstract

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.

Keywords

Metabolic Network Flux Balance Analysis Recombinant Therapeutic Protein Lactate Production Rate Intracellular Metabolite Concentration 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • K.R. de la Luz-Hernández
    • 1
    • 2
  • Y. Rabasa-Legón
    • 1
  • A. Lage-Castellanos
    • 3
  • A. Castillo-Vitlloch
    • 1
  • L. Castellanos-Serra
    • 4
  • J. Díaz-Brito
    • 5
  • S. Gaskell
    • 6
  1. 1.Research and Development DirectionCenter of Molecular ImmunologyHavanaCuba
  2. 2.Michael Barber Center for Mass SpectrometrySchool of Chemistry and Manchester Interdisciplinary Biocenter, University of ManchesterManchesterUK
  3. 3.Cuban Neuroscience CenterHavanaCuba
  4. 4.Center for Genetic Engineering and BiotechnologyHavanaCuba
  5. 5.Faculty of BiologyUniversity of HavanaHavanaCuba
  6. 6.Michael Barber Center for Mass SpectrometrySchool of Chemistry and Manchester Interdisciplinary Biocenter, University of ManchesterManchesterUK

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