Refolding and Purification of rhBMP-2 Expressed as Inclusion Bodies in E.COLI with Hydroxyapatite Chromatography

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The objective of this study was to develop an efficient method for the production of bioactive bone morphogenetic protein 2 (BMP-2). A recombinant plasmid encoding mature human BMP-2 was transferred and expressed at a high level in E.coli. Most of the aimed proteins existed in inclusion bodies. The non-active recombinant human BMP-2 (rhBMP-2) monomer in inclusion bodies was refolded and simultaneously purified using hydroxyapatite (HA) chromatography. After oxidization of the monomer, the rhBMP-2 dimmer showed biological activity by the induction of alkaline phosphate activity in C2C12 cells. The refolding yield was about 30% and the purity was about 90% just by one chromatography process.

Info:

Periodical:

Key Engineering Materials (Volumes 288-289)

Edited by:

Xingdong Zhang, Junzo Tanaka, Yaoting Yu and Yasuhiko Tabata

Pages:

661-664

Citation:

S. Yao et al., "Refolding and Purification of rhBMP-2 Expressed as Inclusion Bodies in E.COLI with Hydroxyapatite Chromatography", Key Engineering Materials, Vols. 288-289, pp. 661-664, 2005

Online since:

June 2005

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$38.00

[1] Rainer Ruppert, Elke Hoffmann, Walter Sebald. Eur. J. Biochem. 1996. 237: 295.

[2] Wang EA. Rosen V. Cordes P etal. Proc. Natl. Acad. Sci. USA. 1988. 85: 9484.

[3] Wang EA. Rosen V. D'Alessandro etal. Proc. Natl. Acad. Sci. USA. 1990. 87: 2220.

[4] Xiong Shaohu, Yu Lei, Tan Haiyan etal. J. First Mil Med. Univ. 2002. 22(5): 413: 416.

[5] Eliana De Bernordez Clark. Current Opinion in Biotechnology. 1998. 9: 157.

[6] Zhenyu Gu. Zhiguo Su. Jan-Christer Janson. J. Chromatogr. A. 2001. 918: 311.

[7] Ming Li. Guifeng Zhang. Zhiguo Su. J. chromatogr. A. 2002 959: 113-120.

[8] Guo LA, Geng XD. Chinese Journal of Biotechnology. 2000. 16(6): 661.

[9] G. Lemercier, N. Bakalara, X. Santarelli. J. Chromatogr. B. 2003. 786: 305.

[10] Cerletti,N. Patent No. US. 6057430.

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