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《China Biotechnology》 2009-08
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Study on pH-responsive Polymer-assisted TGF-β1 Refolding

HUANG Zhi-feng1,2,3 YANG Shu-lin1 NI Chun-yan2 ZHANG Yi2 ZHOU Xian-tian2 LI Xiao-kun1,2(1 School of Chemical Engineering,Nanjing University of Science and Technology,Nanjing 210094,China)(2 Key Laboratory of Biotechnology and Pharmaceutical Engineering of Zhejiang Province,Wenzhou Medical College,Wenzhou 325035,China)(3 School of Chemical and Biomedical Engineering,Nanyang Technological University,Singapore 637459,Singapore)  
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 increased TGF-β1 refolding yield to 53%,when dilution refolding was conducted at 0.5 mg/ml TGF-β1.This shows the evidence of an electrostatic interaction between oppositely charged TGF-β1 and the Eudragit polymer during refolding.This ionic complex of Eudragit and TGF-β1 appears to shield exposed hydrophobic residues of the lysozyme refolding intermediates,thus minimizing hydrophobic-driven aggregation of the molecules.
【Fund】: 国家“863”计划(2007AA02Z110);; 温州市科技攻关项目(Y2007A106)资助项目
【CateGory Index】: Q504
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【Citations】
Chinese Journal Full-text Database 1 Hits
1 LI Xiao-kun, WU Xiao-ping, ZHENG Qing, HUANG Ya-dong,XU Hua, YAO Cheng-can (Biopharmaceutical Research & Development Center,Jinan University,Guangzhou 510632,China);Expression and purification of human transforming growth factor β1 in E.coli RR1[J];Journal of Jinan University(Natural Science & Medicine Edition);2001-05
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