High‐throughput protein refolding screening method using zeolite |
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| Authors: | Takayuki Y Nara Hideaki Togashi Chisato Sekikawa Kengo Sakaguchi Fujio Mizukami Tatsuo Tsunoda |
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| Institution: | 1. Research Center for Compact Chemical Process, AIST, Central 5, 1‐1‐1 Higashi, Tsukuba, Ibaraki 305‐8565, Japan;2. Dept. of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, Noda, Chiba 278‐8510, Japan |
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| Abstract: | We established a 96‐well‐plate‐based refolding screening system using zeolite. In this system, protein denatured and solubilized with 6 M guanidine hydrochloride is adsorbed onto zeolite placed in a 96‐well plate. The refolding conditions can be tested by incubating the samples with refolding buffers under various conditions of pH, salts, and additives. In this study, we chose green fluorescent protein as the model protein. Green fluorescent protein was expressed as inclusion bodies, and we tested the effects of four pH conditions and six additives on its refolding. The results demonstrate that green fluorescent protein was more efficiently refolded with zeolite than with the conventional dilution method. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 |
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| Keywords: | inclusion bodies zeolite refolding screening 96‐well plate green fluorescent protein |
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