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Applied Microbiology and Biotechnology

, Volume 99, Issue 20, pp 8429–8440 | Cite as

Understanding the intracellular effects of yeast extract on the enhancement of Fc-fusion protein production in Chinese hamster ovary cell culture

  • Dongdong Hu
  • Yating Sun
  • Xuping Liu
  • Jintao Liu
  • Xintao Zhang
  • Liang Zhao
  • Haibin Wang
  • Wen-Song TanEmail author
  • Li FanEmail author
Biotechnological products and process engineering

Abstract

Yeast extract (YE), as a non-animal source additive for mammalian cell culture medium, has been widely used for manufacturing of therapeutic proteins. In the present study, one particular YE was found to have significantly improved the specific productivity (q p) of Fc-fusion protein in recombinant Chinese hamster ovary (rCHO) cell culture. In order to elucidate the intracellular effects of YE on protein productivity, steps of the target protein synthesis process were investigated to unveil their variations caused by YE addition. Stepwise analysis on Fc-fusion protein synthesis process showed that YE enhanced Fc-fusion protein gene transcription with cell cycle arrest at G1 phase; mammalian target of rapamycin (mTOR) signaling pathway was activated to enhance the translation of Fc-fusion protein, and the block in post-translational steps of Fc-fusion protein was alleviated by YE addition as well. Our results revealed the responses of multiple protein production steps to the addition of YE and provided a practical guidance for the separation and application of active compounds from hydrolysates.

Keywords

Chinese hamster ovary cells Yeast extract Fc-fusion protein Specific productivity Cell cycle arrest mTOR 

Notes

Acknowledgments

This work was supported by the National Natural Science Foundation of China (Nos. 21206040 and 21406066), the National Science and Technology Major Project (No. 2013ZX10004003-003-003), and the National High Technology Research and Development Program of China (863 Program) (No. 2012AA02A303).

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

253_2015_6789_MOESM1_ESM.pdf (112 kb)
ESM 1 (PDF 112 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Dongdong Hu
    • 1
  • Yating Sun
    • 1
  • Xuping Liu
    • 1
  • Jintao Liu
    • 1
  • Xintao Zhang
    • 1
  • Liang Zhao
    • 1
  • Haibin Wang
    • 2
  • Wen-Song Tan
    • 1
    Email author
  • Li Fan
    • 1
    Email author
  1. 1.State Key Laboratory of Bioreactor EngineeringEast China University of Science and TechnologyShanghaiPeople’s Republic of China
  2. 2.Zhejiang Hisun Pharmaceutical Co., Ltd.HangzhouChina

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