Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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High-Level Expression of Recombinant Human Bone Morphogenetic Protein-4 in Chinese Hamster Ovary Cells

  • PARK JUNHO (Institute of Life Sciences and Resources, and Graduate School of Biotechnology, Kyung Hee University) ;
  • YU SUNGRYUL (Institute of Life Sciences and Resources, and Graduate School of Biotechnology, Kyung Hee University) ;
  • YOON JAESEUNG (Institute of Life Sciences and Resources, and Graduate School of Biotechnology, Kyung Hee University) ;
  • BAEK KWANGHEE (Institute of Life Sciences and Resources, and Graduate School of Biotechnology, Kyung Hee University)
  • Published : 2005.12.01
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Abstract

Bone morphogenetic protein-4 (BMP-4) is a signaling homodimeric molecule that acts as a morphogen to influence cell fate in a concentration-dependent manner. The limited supply of a pure preparation of BMP-4, due to very low level of their expression in vivo, makes it difficult not only to study the biological activities of BMPs, but also to use them as a clinical tool. For a large-scale production of BMP-4, human BMP-4 cDNA was expressed in Chinese hamster ovary (CHO) cells by a recently development vector system, which confers position-independent stable expression of the foreign genes. The CHO cell line expressing recombinant human BMP-4 (rhBMP-4) at the level of $7\;{\mu}g/ml$ could be obtained after stepwise selection with methotrexate. This level of expression is about 70 times higher than those previously reported. The partially processed form of BMP-4 as well as mature form could be detected, when the aliquots of culture media were analyzed by Western blot. The glycosylation pattern and biological activity of the rhBMP-4 were determined by glycosidase treatment and the induction rate of alkaline phosphatase in mouse osteoblastic cells.

Keywords

References

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