Artificial chaperone-assisted refolding in a microchannel

作者：Yamamoto Etsushi; Yamaguchi Satoshi^*; Sasaki Naoki; Kim Haeng Boo; Kitamori Takehiko; Nagamune Teruyuki

来源：Bioprocess and Biosystems Engineering, 2010, 33(1): 171-177.

DOI：10.1007/s00449-009-0374-1

• Alpha-glucosidase
• Artificial chaperone
• Beta-cyclodextrin
• Lysozyme
• Microchannel
• Protein refolding

Protein refolding using a simple dilution method in a microchannel often led to the formation of protein aggregates, which bound to the microchannel wall, resulting in low refolding yields. To inhibit aggregation and improve refolding yields, an artificial chaperone-assisted (ACA) refolding, which employed detergents and beta-cyclodextrin was used. Model proteins, hen egg white lysozyme and yeast alpha-glucosidase, were successfully refolded in a microchannel. The microscopic observation showed that the ACA method suppressed protein aggregation and facilitated the refolding of lysozyme, whereas significant aggregation was observed when a simple dilution method was employed. The ACA method increased the lysozyme refolding yield by 40% over the simple dilution approach. Similarly, for alpha-glucosidase, the refolding yield using the ACA method (ca. 50%) was approximately three times compared with the simple dilution method. The ACA refolding method is a suitable approach to use in the refolding of proteins using a microfluidic system.

• 出版日期2010-1