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Study of pH-responsive Polymer-assisted TGF-β1 Refolding |
HUANG Zhi-Feng1,2,3, YANG Shu-Lin1, NI Chun-Yan2, ZHANG Yi2, ZHOU Xiang-Tian2, LI Jiao-Kun1,2 |
1.School of Chemical Engineering, Nanjing University of Science and Technology, Nanjing210094, China
2. Key Laboratory of Biotechnology and Pharmaceutical Engineering of Zhejiang Province, Wenzhou Medical College, Wenzhou325035, China
3.School of Chemical and Biomedical Engineering, Nanyang Technological University,Singapore637459,Singapore |
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Abstract In the present work we have studied if Eudragit S-100, a pH-responsive polymer can enhance the refolding level of TGF-β1 (Transforwing growth factor,TGF-β1). The refolding of TGF-β1 was performed by directly diluting denatured TGF-β1 into a refolding buffer containing different concentration of Eudragit. The ability of Eudragit S-100 to enhance protein refolding level was investigated using MTT assay, reverse phase HPLC, fluorescence emission spectroscopy and circular dichroism spectroscopy. The addition of Eudragit S-100 in the refolding buffer significantly increases TGF-β1 refolding yield to 53%, when dilution refolding was conducted at 0.5 mg/ml TGF-β1. This study shows evidence of an electrostatic interaction between oppositely charged TGF-β1 and the Eudragit polymer during refolding. This ionic complexing of Eudragit and TGF-β1 appears to shield exposed hydrophobic residues of the lysozyme refolding intermediates, thus minimizing hydrophobic-driven aggregation of the molecules.
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Received: 12 March 2009
Published: 28 July 2009
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