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KMID : 0380020030180060500
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Korean Journal of Biotechnology and Bioengineering
2003 Volume.18 No. 6 p.500 ~ p.505
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Solid-Phase Refolding of Inclusion Body Protein in Packed Bed Adsorption and Expanded Bed Adsorption Chromatography
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Choi Won-Chan
Kim Min-Young
Seo Chang-Woo
Lee Eun-Kyu
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Abstract
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‘LK (lipoprotein kringle) 68’is a polypeptide of a modified ansiostatin consisting of three kringle structures that might be clinically useful as a potential cancer therapeutics. It can be produced by overexpressing it as inclusion body in recombinant E. coli. In this study, solid-phase refolding processes using packed bed adsorption (PBA) and expanded bed adsorption (EBA) column were carried out to compare their refolding yields with that of the conventional, solution-phase refolding process, For the solution-phase and the PBA-mediated processes employing Q-Sepharose, washed inclusion body was used as the starting material, whereas both washed inclusion body and E. coli homogenate were used for the EBA-mediated process employing streamline DEAE. On the final recovery LK68 per unit mass of wet cell basis, the EBA- and PBA-mediated processes showed about 2.7- and 1.5-fold higher yields, respectively, than the solution-phase refolding method. The solid-phase refolded LK68 demonstrated the same Iysine binding bioactivity and the retention time in the RP-and SEC-HPLC as those of the native protein.
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KEYWORD
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inclusion body, solid-phase refolding, renaturation, angiostatin, expanded bed adsorption chromatography
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