期刊
BIOTECHNOLOGY AND BIOENGINEERING
卷 109, 期 6, 页码 1395-1403出版社
WILEY
DOI: 10.1002/bit.24442
关键词
rCHO cells; glycine betaine; hyperosmolality; proteomics; DIGE
资金
- MEST [R31-2008-000-10071-0]
- Ministry of Knowledge Economy
- WCU through the KOSEF
- 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)
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 (beta-tubulin and beta-actin), protein folding (GRP78 and OSP94), mRNA processing (Rbm34, ACF, and IPMK), and protein secretion (?-COP). 2D-Western blot analysis of beta-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; 109:13951403. (c) 2012 Wiley Periodicals, Inc.
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