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Differential in-gel electrophoresis (DIGE) analysis of CHO cells under hyperosmotic pressure: Osmoprotective effect of glycine betaine addition

Authors

  • Jee Yon Kim,

    1. Department of Biological Sciences, Graduate School of Nanoscience and Technology (WCU), KAIST, 373-1 Kusong-Dong, Yusong-Gu, Daejon 305-701, Korea; telephone: +82-42-350-2618; fax: +82-42-350-2610
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  • Yeon-Gu Kim,

    1. Biotechnology Process Engineering Center, KRIBB, Daejeon, Korea
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  • Gyun Min Lee

    Corresponding author
    1. Department of Biological Sciences, Graduate School of Nanoscience and Technology (WCU), KAIST, 373-1 Kusong-Dong, Yusong-Gu, Daejon 305-701, Korea; telephone: +82-42-350-2618; fax: +82-42-350-2610
    • Department of Biological Sciences, Graduate School of Nanoscience and Technology (WCU), KAIST, 373-1 Kusong-Dong, Yusong-Gu, Daejon 305-701, Korea; telephone: +82-42-350-2618; fax: +82-42-350-2610.
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  • Jee Yon Kim and Yeon-Gu Kim contributed equally to this work.

Abstract

The use of glycine betaine combined with hyperosmolality is known to be an efficient means for achieving high protein production in recombinant Chinese hamster ovary (rCHO) cells. In order to understand the intracellular events and identify the key factors in rCHO cells cultivated with glycine betaine under hyperosmotic conditions, two-dimensional differential in-gel electrophoresis (2D-DIGE) followed by mass spectrometric analysis was applied. Differentially expressed 19 protein spots were selected and 16 different kinds of proteins were successfully identified. The identified proteins were associated with cellular metabolism (PEPCK, GAPDH, and PK), cellular architecture (β-tubulin and β-actin), protein folding (GRP78 and OSP94), mRNA processing (Rbm34, ACF, and IPMK), and protein secretion (γ-COP). 2D-Western blot analysis of β-tubulin, GAPDH, Peroxidoxin-1, and GRP78 confirmed the proteomic findings. The proteins identified from this study, which are related to cell growth and antibody production, can be applied to cell engineering for maximizing the efficacy of the use of glycine betaine combined with hyperosmolality in rCHO cells. Biotechnol. Bioeng. 2012; 109:1395–1403. © 2012 Wiley Periodicals, Inc.

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