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Proteomic understanding of intracellular responses of recombinant Chinese hamster ovary cells cultivated in serum-free medium supplemented with hydrolysates

  • Genomics, Transcriptomics, Proteomics
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

In order to understand the intracellular responses in recombinant CHO (rCHO) cells producing antibody in serum-free medium (SFM) supplemented with optimized hydrolysates mixtures, yielding the highest specific growth rate (μ, SFM#S1) or the highest specific antibody productivity (q Ab, SFM#S2), differentially expressed proteins in rCHO cells are measured by two-dimensional gel electrophoresis combined with nano-LC-ESI-Q-TOF tandem MS. The comparative proteomic analysis with basal SFM without hydrolysates revealed that the addition of hydrolysate mixtures significantly altered the profiles of CHO proteome. In SFM#S1, the expression of metabolism-related proteins, cytoskeleton-associated proteins, and proliferation-related proteins was up-regulated. On the other hand, the expression of anti-proliferative proteins and pro-apoptotic protein was down-regulated. In SFM#S2, the expression of various chaperone proteins and proliferation-linked proteins was altered. 2D-Western blot analysis of differentially expressed proteins confirmed the proteomic results. Taken together, identification of differentially expressed proteins in CHO cells by a proteomic approach can provide insights into understanding the effect of hydrolysates on intracellular events and clues to find candidate genes for cell engineering to maximize the protein production in rCHO cells.

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Acknowledgments

This research was supported in part by WCU program through the KOSEF funded by the MEST (grant number: R31-2008-000-10071-0) and the Ministry of Knowledge Economy (Strategic Technology Development Project). YH Kim was supported by a grant of Korea Basic Science Institute (G30123).

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Authors and Affiliations

  1. Department of Biological Sciences and Graduate School of Nanoscience & Technology (WCU), KAIST, 373-1 Kusong-Dong, Yuseong-Gu, Daejon, 305-701, South Korea

    Jee Yon Kim, Yeon-Gu Kim, Young Kue Han & Gyun Min Lee

  2. Division of Mass Spectrometry Research, Korea Basic Science Institute, 804-1 Yangcheong-Ri, Ochang-Eup, Cheongwon-Gun, Chungbuk, 863-883, South Korea

    Hyun Soo Choi & Young Hwan Kim

  3. Graduate School of Analytical Science and Technology, Chungnam National University, 79 Daehangno, Yuseong-Gu, Daejeon, 305-764, South Korea

    Young Hwan Kim

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  1. Jee Yon Kim
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  2. Yeon-Gu Kim
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  3. Young Kue Han
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  4. Hyun Soo Choi
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Correspondence to Young Hwan Kim or Gyun Min Lee.

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Kim, J.Y., Kim, YG., Han, Y.K. et al. Proteomic understanding of intracellular responses of recombinant Chinese hamster ovary cells cultivated in serum-free medium supplemented with hydrolysates. Appl Microbiol Biotechnol 89, 1917–1928 (2011). https://doi.org/10.1007/s00253-011-3106-9

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  • DOI: https://doi.org/10.1007/s00253-011-3106-9

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