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On‐column refolding of corticotropin‐releasing factor receptor 1 extracellular domain by size exclusion chromatography
Author(s) -
Ersoy Maria,
Matic Sandra,
Schmidt Stefan R.
Publication year - 2009
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1233
Subject(s) - chemistry , chromatography , size exclusion chromatography , guanidine , extracellular , inclusion bodies , column chromatography , affinity chromatography , escherichia coli , biochemistry , enzyme , gene
Expressing the extracellular domain of corticotropin releasing factor receptor 1 in Escherichia coli usually results in the formation of inclusion bodies. Here we describe the optimization of refolding by applying size exclusion chromatography with a denaturing guanidine hydrochloride gradient and a refolding buffer containing glycerol. Several chromatographic parameters like gradient length, flow rate, sample concentration and chromatography resin characteristics were evaluated. Recovery yields of refolded protein above 50% using a Superdex 200 column demonstrate the usefulness of this method. Copyright © 2009 John Wiley & Sons, Ltd.