z-logo
Premium
On‐Column Refolding of Recombinant Human Interferon‐γ with an Immobilized Chaperone Fragment
Author(s) -
Gao YongGui,
Guan YiXin,
Yao ShanJing,
Cho ManGi
Publication year - 2003
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp025775l
Subject(s) - groel , chemistry , sephadex , chaperone (clinical) , chromatography , recombinant dna , inclusion bodies , protein folding , specific activity , in vitro , biochemistry , biophysics , escherichia coli , enzyme , biology , medicine , pathology , gene
The chaperone mini‐GroEL is a soluble recombinant fragment containing the 191–345 amino acid sequence of GroEL with a 6×His tag. The refolding protocol assisted with mini‐GroEL was studied for the activity recovery of rhIFN‐γ inclusion bodies. In a suspended system, mini‐GroEL showed significant enhancement of the activity recovery of rhIFN‐γ, applyed with a 1–5:1 stoichiometry of mini‐GroEL to rhIFN‐γ at 25 °C. Moreover, 1 M urea in the renaturation buffer had a synergistic effect on suppressing the aggregation and improving the activity recovery. Finally, a novel chromatographic column, containing 1 cm height of Sephadex G 200 at the top of column and packed with immobilized mini‐GroEL to promote refolding, was devised. The total activity recovered per milligram of denatured rhIFN‐γ was up to 3.93 × 10 6 IU with the immobilized mini‐GroEL column, which was reused four times without evident loss of renaturation ability. A convenient technique with the integrated process of chaperon preparation and rhIFN‐γ folding in vitro was developed.

This content is not available in your region!

Continue researching here.

Having issues? You can contact us here