Artificial chaperone-assisted refolding in a microchannel |
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| Authors: | Etsushi Yamamoto Satoshi Yamaguchi Naoki Sasaki Haeng-Boo Kim Takehiko Kitamori Teruyuki Nagamune |
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| Institution: | (1) Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan;(2) Department of Bioengineering, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan;(3) Center for NanoBio Integration (CNBI), The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan;(4) Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan;(5) Microchemistry Group, Kanagawa Academy of Science and Technology (KAST), Ksp Bldg. East 307, 3-2-1 Sakado, Takahatsu-ku, Kawasaki 213-0012, Japan;(6) Present address: Bioengineering Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan;(7) Present address: Faculty of Science, Ibaraki University, Bunkyo 2-1-1 Mito, Ibaraki 310-8512, Japan; |
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| Abstract: | 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 β-cyclodextrin was used. Model proteins, hen
egg white lysozyme and yeast α-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 α-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. |
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