Nano-cage-mediated refolding of insulin by PEG-PE micelle

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• 作者：Xiaocui Fang^a ; ^b ; ¹ ; Tao Yang^a ; ^c ; ¹ ; Luoyang Wang^a ; ^c ; Jibing Yu^a ; Xiuli Wei^a ; Yinjian Zhou^a ; Chen Wang^b ; ^{wangch@nanoctr.cn" class="auth_mail" title="E-mail the corresponding author} ; Wei Liang^a ; ^{weixx@sun5.ibp.ac.cn" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Insulin ; DTT ; Refolding ; PEG-PE micelle ; Nano-cage ; Negative charge layer
• 刊名：Biomaterials
• 出版年：2016
• 出版时间：January 2016
• 年：2016
• 卷：77
• 期：Complete
• 页码：139-148
• 全文大小：2586 K

文摘

Insulin aggregation has pronounced pharmaceutical implications and biological importance. Deposition of insulin aggregates is associated with type II diabetes and instability of pharmaceutical formulations. We present in this study the renaturation effect of PEG-PE micelle on dithiothreitol (DTT)-denatured insulin revealed by techniques including turbidity assay, circular dichroism (CD), thioflavinT (ThT) binding assay, bis-ANS binding assay, agarose gel electrophoresis and MALDI-TOF MS. The obtained results show that PEG-PE micelle having a hydrophilic nano-cage-like structure in which with a negative charge layer, can capture DTT-induced insulin A and B chains, and block their hydrophobic interaction, thereby preventing aggregation. The reduced insulin A and B chain in the nano-cage are capable of recognizing each other and form the native insulin with yields of ∼30% as measured by hypoglycemic activity analysis in mice. The observed insulin refolding assisted by PEG-PE micelle may be applicable to other proteins.