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HUANG Zhi-feng, YANG Shu-lin, ZHANG Yi, FENG Wen-ke, LI Xiao-kun. Assistance of Eudragit S-100 on protein refolding[J]. Journal of China Pharmaceutical University, 2009, 40(3): 273-278.
Citation: HUANG Zhi-feng, YANG Shu-lin, ZHANG Yi, FENG Wen-ke, LI Xiao-kun. Assistance of Eudragit S-100 on protein refolding[J]. Journal of China Pharmaceutical University, 2009, 40(3): 273-278.

Assistance of Eudragit S-100 on protein refolding

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  • Aim :To study the effect of Eudragit S-100,a pH-responsive polymer,on protein refolding level, using recombinant human keratinocyte growth factor-2(rhKGF-2) as a model protein. Methods :The refolding of rhKGF-2 was performed by directly diluting denatured rhKGF-2 into a refolding buffer containing different concentrations 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. Results :The addition of Eudragit S-100 in the refolding buffer significantly increased the rhKGF-2 refolding yield to 71%,when dilution refolding was conducted at 0.5 mg/mL rhKGF-2.The outcome from the refolding study showed possibility of a special interaction between rhKGF-2 and Eudragit,suggesting that the refolding-enhancing ability of Eudragit S-100 was due to this interaction between Eudragit S-100 and rhKGF-2.Mean while,the result showed that the concentration of urea was also an important factor for the optimization of the refolding in the presence of Eudragit. Conclusion :Eudragit S-100 can significantly increase the refolding level of rhKGF-2.
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