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J. Microbiol. Biotechnol. 2006; 16(5): 695-703

Published online May 28, 2006

Copyright © The Korean Society for Microbiology and Biotechnology.

High Productivity of t-PA in CHO Cells Using Hypoxia Response Element

Hong Jin Kim , Hongwoo Park 1, Seongman Kang 2, Bae, Gun Won 2, Dae-Won Jeong 3, Gyun Min Lee 4, Tae-Boo Choe 5, Ick Young Kim 5 and Ik-Hwan Kim 5*

Microbiology Section, College of Pharmacy, Chung Ang University, Seoul 156-756, Korea, 1Division of Applied Chemical Engineering & Bio Engineering, Seoul 133-791, Korea, 2College of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Korea, 3BK21 HLS, Seoul National University, Seoul 110-749, Korea, 4Department of Biological Sciences, KAIST, Daejeon 305-701, Korea, 5Department of Microbiological Engineering, Seoul 143-701, Korea

Abstract

The dissolved oxygen level of any cell culture environment has a critical effect on cellular metabolism. Specifically, hypoxia condition decreases cell viability and recombinant protein productivity. In this work, to develop CHO cells producing recombinant protein with high productivity, mammalian expression vectors containing a human tissue-type plasminogen activator (t-PA) gene with hypoxia response element (HRE) were constructed and stably transfected into CHO cells. CHO/2HRE-t-PA cells produced 2-folds higher recombinant t-PA production than CHO/t-PA cells in a $Ba^{2+}-alginate$ immobilized culture, and 16.8-folds in a repeated batch culture. In a non-aerated batch culture of suspension-adapted cells, t-PA productivity of CHO/2HRE/t-PA cells was 4.2-folds higher than that of CHO/t-PA cells. Our results indicate that HRE is a useful tool for the enhancement of protein productivity in mammalian cell cultures.

Keywords: Chinese hamster ovary cell, hypoxia, inducible factor-1α, tissue-type plasminogen activator, hypoxia inducible factor-1α, immobilized cells