Premium
Differential in‐gel electrophoresis (DIGE) analysis of CHO cells under hyperosmotic pressure: Osmoprotective effect of glycine betaine addition
Author(s) -
Kim Jee Yon,
Kim YeonGu,
Lee Gyun Min
Publication year - 2012
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.24442
Subject(s) - betaine , chinese hamster ovary cell , glycine , biochemistry , western blot , gel electrophoresis , glyceraldehyde , chemistry , proteome , osmotic concentration , biology , microbiology and biotechnology , enzyme , amino acid , gene , dehydrogenase , receptor
The use of glycine betaine combined with hyperosmolality is known to be an efficient means for achieving high protein production in recombinant Chinese hamster ovary (rCHO) cells. In order to understand the intracellular events and identify the key factors in rCHO cells cultivated with glycine betaine under hyperosmotic conditions, two‐dimensional differential in‐gel electrophoresis (2D‐DIGE) followed by mass spectrometric analysis was applied. Differentially expressed 19 protein spots were selected and 16 different kinds of proteins were successfully identified. The identified proteins were associated with cellular metabolism (PEPCK, GAPDH, and PK), cellular architecture (β‐tubulin and β‐actin), protein folding (GRP78 and OSP94), mRNA processing (Rbm34, ACF, and IPMK), and protein secretion (γ‐COP). 2D‐Western blot analysis of β‐tubulin, GAPDH, Peroxidoxin‐1, and GRP78 confirmed the proteomic findings. The proteins identified from this study, which are related to cell growth and antibody production, can be applied to cell engineering for maximizing the efficacy of the use of glycine betaine combined with hyperosmolality in rCHO cells. Biotechnol. Bioeng. 2012; 109:1395–1403. © 2012 Wiley Periodicals, Inc.