High recovery refolding of rhG-CSF from Escherichia coli, using urea gradient size exclusion chromatography

Biotechnology Progress
Chaozhan WangXindu Geng

Abstract

Protein folding liquid chromatography (PFLC) is a powerful tool for simultaneous refolding and purification of recombinant proteins in inclusion bodies. Urea gradient size exclusion chromatography (SEC) is a recently developed protein refolding method based on the SEC refolding principle. In the presented work, recombinant human granulocyte colony-stimulating factor (rhG-CSF) expressed in Escheriachia coli (E. coli) in the form of inclusion bodies was refolded with high yields by this method. Denatured/reduced rhG-CSF in 8.0 mol.L(-1) urea was directly injected into a Superdex 75 column, and with the running of the linear urea concentration program, urea concentration in the mobile phase and around the denatured rhG-CSF molecules was decreased linearly, and the denatured rhG-CSF was gradually refolded into its native state. Aggregates were greatly suppressed and rhG-CSF was also partially purified during the refolding process. Effects of the length and the final urea concentration of the urea gradient on the refolding yield of rhG-CSF by using urea gradient SEC were investigated respectively. Compared with dilution refolding and normal SEC with a fixed urea concentration in the mobile phase, urea gradient SEC was more efficient...Continue Reading

Citations

May 3, 2008·Journal of Proteome Research·Tasneem H PatwaDavid M Lubman
Jun 12, 2013·Protein Expression and Purification·Anuja M RaneZhiyu Li
Jun 16, 2009·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·K K FursovaF A Brovko
Nov 1, 2012·Journal of Separation Science·Fei WangXindu Geng
Oct 23, 2012·Journal of Separation Science·Pratap D BadeAnurag S Rathore
Mar 27, 2021·Frontiers in Bioengineering and Biotechnology·Sonal Datta

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