On‐column refolding of corticotropin‐releasing factor receptor 1 extracellular domain by size exclusion chromatography | Zendy

Ersoy Maria | Zendy; Matic Sandra | Zendy; Schmidt Stefan R. | Zendy

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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.

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