Genomics, Transcriptomics, Proteomics

Applied Microbiology and Biotechnology

, Volume 89, Issue 6, pp 1917-1928

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

  • Jee Yon KimAffiliated withDepartment of Biological Sciences and Graduate School of Nanoscience & Technology (WCU)
  • , Yeon-Gu KimAffiliated withDepartment of Biological Sciences and Graduate School of Nanoscience & Technology (WCU)
  • , Young Kue HanAffiliated withDepartment of Biological Sciences and Graduate School of Nanoscience & Technology (WCU)
  • , Hyun Soo ChoiAffiliated withDivision of Mass Spectrometry Research, Korea Basic Science Institute
  • , Young Hwan KimAffiliated withDivision of Mass Spectrometry Research, Korea Basic Science InstituteGraduate School of Analytical Science and Technology, Chungnam National University Email author 
  • , Gyun Min LeeAffiliated withDepartment of Biological Sciences and Graduate School of Nanoscience & Technology (WCU) Email author 

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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.

Keywords

CHO cells Proteomics Hydrolysates Cell growth Productivity