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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제27권 제7호
- 발행연도
- 2017.7
- 수록면
- 1,281 - 1,287 (7page)
- DOI
- 10.4014/jmb.1702.02011
이용수
초록· 키워드
오류제보하기
Bone morphogenetic protein-4 (BMP-4) is considered to have therapeutic potential for various diseases, including cancers; however, the high expression of biologically active recombinant human BMP-4 (rhBMP-4) needed for its manufacture for therapeutic purposes has yet to be established. In the current study, we established a recombinant Chinese hamster ovary (rCHO) cell line overexpressing rhBMP-4 as well as a production process using 7.5-l bioreactor (5 L working volume). The expression of the mature rhBMP-4 was significantly enhanced by recombinant furin expression. The combination of a chemically defined medium and a nutrient supplement solution for high expression of rhBMP-4 was selected and used for bioreactor cultures. The 11-day fed-batch cultures of the established rhBMP-4-expressing rCHO cells in the 7.5-L bioreactor produced approximately 32 mg/l of rhBMP-4. The mature rhBMP-4 was purified to homogeneity from the culture supernatant using a two-step chromatographic procedure, resulting in a recovery rate of approximately 55% and a protein purity greater than 95%. The N-terminal amino acid sequences and N-linked glycosylation of the purified rhBMP-4 were confirmed by N-terminal sequencing and de-N-glycosylation analysis, respectively. The mature purified rhBMP-4 has been proved to be functionally active, with an effective dose concentration of EC50 of 2.93 ng/ml.
목차
등록된 정보가 없습니다.
참고문헌 (25)
참고문헌 신청
Celeste AJ / 1990 / Identification of transforming growth factor beta family members present in bone-inductive protein purified from bovine bone / Proc. Natl. Acad. Sci. USA 87:9843~9847
Wozney JM / 1988 / Novel regulators of bone formation:molecular clones and activities / Science 242:1528~1534
Miyazaki Y / 2000 / Bone morphogenetic protein 4 regulates the budding site and elongation of the mouse ureter / J. Clin. Invest 105:863~873
Winnier G / 1995 / Bone morphogenetic protein-4 is required for mesoderm formation and patterning in the mouse / Genes Dev 9:2105~2116
Bragdon B / 2011 / Bone morphogenetic proteins: a critical review / Cell Signal 23:609~620
Wozney JM / 1988 / Novel regulators of bone formation:molecular clones and activities / Science 242:1528~1534
Miyazaki Y / 2000 / Bone morphogenetic protein 4 regulates the budding site and elongation of the mouse ureter / J. Clin. Invest 105:863~873
Winnier G / 1995 / Bone morphogenetic protein-4 is required for mesoderm formation and patterning in the mouse / Genes Dev 9:2105~2116
Bragdon B / 2011 / Bone morphogenetic proteins: a critical review / Cell Signal 23:609~620
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