Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 89, Issue 6, Pages 1917-1928Publisher
SPRINGER
DOI: 10.1007/s00253-011-3106-9
Keywords
CHO cells; Proteomics; Hydrolysates; Cell growth; Productivity
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Funding
- MEST [R31-2008-000-10071-0]
- Ministry of Knowledge Economy
- Korea Basic Science Institute [G30123]
- Ministry of Education, Science & Technology (MoST), Republic of Korea [R31-2008-000-10071-0] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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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 (mu, 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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