title: Understanding the intracellular effects of yeast extract on the enhancement of Fc-fusion protein production in Chinese hamster ovary cell culture.
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.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, P. O. Box 309, 130 Mei-Long Road, Shanghai, 200237, People's Republic of China.
authors.forename: Dongdong
authors.initials: D
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, P. O. Box 309, 130 Mei-Long Road, Shanghai, 200237, People's Republic of China.
authors.forename: Yating
authors.initials: Y
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, P. O. Box 309, 130 Mei-Long Road, Shanghai, 200237, People's Republic of China.
authors.forename: Xuping
authors.initials: X
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, P. O. Box 309, 130 Mei-Long Road, Shanghai, 200237, People's Republic of China.
authors.forename: Jintao
authors.initials: J
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, P. O. Box 309, 130 Mei-Long Road, Shanghai, 200237, People's Republic of China.
authors.forename: Xintao
authors.initials: X
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, P. O. Box 309, 130 Mei-Long Road, Shanghai, 200237, People's Republic of China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Zhejiang Hisun Pharmaceutical Co., Ltd., Fuyang, Hangzhou, Zhejiang, 311404, China.
authors.forename: Haibin
authors.initials: H
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, P. O. Box 309, 130 Mei-Long Road, Shanghai, 200237, People's Republic of China. wstan@ecust.edu.cn.
authors.forename: Wen-Song
authors.initials: WS
authors.lastname: Tan
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, P. O. Box 309, 130 Mei-Long Road, Shanghai, 200237, People's Republic of China. fanli@ecust.edu.cn.
authors.forename: Li
authors.initials: L
authors.lastname: Fan
mesh_terms: D000818:Animals; D016466:CHO Cells / Q000378:metabolism*; D018929:Cell Culture Techniques; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000378:metabolism*; D010461:Peptones / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D003470:Culture Media; D007141:Immunoglobulin Fc Fragments; D010461:Peptones; D011993:Recombinant Fusion Proteins
keywords: Cell cycle arrest; Chinese hamster ovary cells; Fc-fusion protein; Specific productivity; Yeast extract; mTOR