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Title 

Differential in-gel electrophoresis (DIGE) analysis of CHO cells under hyperosmotic pressure: Osmoprotective effect of glycine betaine addition

Authors 

J Y KimYeon Gu KimG M Lee

Publisher 

Wiley-Blackwell

Issue Date 

2012

Citation 

Biotechnology and Bioengineering, vol. 109, no. 6, pp. 1395-1403

Keywords 

DIGEGlycine betaineHyperosmolalityProteomicsRCHO cells

Abstract 

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.

ISSN 

0006-3592

Link 

http://dx.doi.org/10.1002/bit.24442

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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