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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2017.1
- 수록면
- 71 - 78 (8page)
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초록· 키워드
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BMP-2 is a well-known TGF-beta related growth factor, having a significant role in bone and cartilage formation. It has been employed to promote bone formation in some clinical trials, and to differentiate mesenchymal stem cells into osteoblasts. However, it is difficult to obtain this protein in its soluble and active form. hBMP-2 is expressed as an inclusion body in the bacterial system. To continuously supply hBMP-2 for research, we optimized the refolding of recombinant hBMP-2 expressed in E. coli, and established an efficient method by using detergent and alkali. Using a heparin column, the recombinant hBMP-2 was purified with the correct refolding. Although combinatorial refolding remarkably enhanced the solubility of the inclusion body, a higher yield of active dimer form of hBMP-2 was obtained from one-step refolding with detergent. The refolded recombinant hBMP-2 induced alkaline phosphatase activity in mouse myoblasts, at ED50 of 300-480ng/ml. Furthermore, the expressions of osteogenic markers were upregulated in hPDLSCs and hDPSCs.
Therefore, using the process described in this study, the refolded hBMP-2 might be cost-effectively useful for various differentiation experiments in a laboratory.
목차
등록된 정보가 없습니다.
참고문헌 (31)
참고문헌 신청
Surinder Mohan Singh / 2005 / Solubilization and refolding of bacterial inclusion body proteins / Journal of Bioscience and Bioengineering 99(4):303~310
St John RJ / 1999 / High pressure fosters protein refolding from aggregates at high concentrations / Proc Natl Acad Sci U S A 96:13029~13033
Burgess RR / 1996 / Purification of overproduced Escherichia coli RNA polymerase sigma factors by solubilizing inclusion bodies and refolding from Sarkosyl / Methods Enzymol 273:145~149
Motonori Kudou / 2011 / Refolding single-chain antibody (scFv) using lauroyl-l-glutamate as a solubilization detergent and arginine as a refolding additive / Protein Expression and Purification 77(1):68~74
Surinder M. Singh / 2012 / Solubilization of inclusion body proteins using n -propanol and its refolding into bioactive form / Protein Expression and Purification 81(1):75~82
St John RJ / 1999 / High pressure fosters protein refolding from aggregates at high concentrations / Proc Natl Acad Sci U S A 96:13029~13033
Burgess RR / 1996 / Purification of overproduced Escherichia coli RNA polymerase sigma factors by solubilizing inclusion bodies and refolding from Sarkosyl / Methods Enzymol 273:145~149
Motonori Kudou / 2011 / Refolding single-chain antibody (scFv) using lauroyl-l-glutamate as a solubilization detergent and arginine as a refolding additive / Protein Expression and Purification 77(1):68~74
Surinder M. Singh / 2012 / Solubilization of inclusion body proteins using n -propanol and its refolding into bioactive form / Protein Expression and Purification 81(1):75~82
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