Understanding the intracellular effects of yeast extract on the enhancement of Fc-fusion protein production in Chinese hamster ovary cell culture
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.
KeywordsChinese hamster ovary cells Yeast extract Fc-fusion protein Specific productivity Cell cycle arrest mTOR
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.
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