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

Genomics, Transcriptomics, Proteomics

DOI: 10.1007/s00253-011-3106-9

Cite this article as:
Kim, J.Y., Kim, YG., Han, Y.K. et al. Appl Microbiol Biotechnol (2011) 89: 1917. doi:10.1007/s00253-011-3106-9

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 (qAb, 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 cellsProteomicsHydrolysatesCell growthProductivity

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Department of Biological Sciences and Graduate School of Nanoscience & Technology (WCU)DaejonSouth Korea
  2. 2.Division of Mass Spectrometry Research, Korea Basic Science InstituteCheongwon-GunSouth Korea
  3. 3.Graduate School of Analytical Science and TechnologyChungnam National UniversityYuseong-GuSouth Korea