Urea gradient size-exclusion chromatography enhanced the yield of lysozyme refolding
Department of Biochemistry and Molecular Pharmacology
Chromatography, Gel; Chromatography, Ion Exchange; Muramidase; *Protein Folding; Urea
Biochemistry, Biophysics, and Structural Biology | Pharmacology, Toxicology and Environmental Health
Protein refolding is still a bottleneck for large-scale production of valuable proteins expressed as inclusion bodies in Escherichia coli. Usually biologically active proteins cannot be obtained with high yield at a high concentration after refolding. In order to meet the challenge of protein refolding a urea gradient gel filtration-refolding system was developed in this article. A Superdex 75 column was pre-equilibrated with a linear decreased urea gradient, the denatured protein experienced the gradual decrease in urea concentration as it went through the column. The refolding of denatured lysozyme showed this method could significantly increase the activity recovery of denatured lysozyme at high protein concentration. The activity recovery of 90% was obtained from the initial protein concentration up to 17 mg/ml within 40 min.
J Chromatogr A. 2001 May 25;918(2):311-8.
Journal of chromatography. A
Gu Z, Su Z, Janson J. (2001). Urea gradient size-exclusion chromatography enhanced the yield of lysozyme refolding. Biochemistry and Molecular Pharmacology Publications. Retrieved from https://escholarship.umassmed.edu/bmp_pp/120