Improving the refolding of NTA protein by urea gradient and arginine gradient size-exclusion chromatography
| Autor: | Xiangdong Fan, DianSheng Xu, Bing Lu, Jie Xia, Dong-Zhi Wei |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Protein Denaturation
Protein Folding Chromatography Arginine High protein Protein Renaturation Size-exclusion chromatography Biophysics Biochemistry Recombinant Proteins law.invention chemistry.chemical_compound Fibrinolytic Agents chemistry law Chromatography Gel Escherichia coli Recombinant DNA Urea Muramidase Protein activity Lysozyme Protein concentration |
| Zdroj: | Journal of Biochemical and Biophysical Methods. 70:1130-1138 |
| ISSN: | 0165-022X |
| DOI: | 10.1016/j.jprot.2007.12.004 |
| Popis: | Inclusion body refolding processes play a major role in the production of recombinant proteins. Improvement of the size-exclusion chromatography refolding process was achieved by combining a decreasing urea gradient with an increasing arginine gradient (two gradients) for the refolding of NTA protein (a new thrombolytic agent) in this paper. Different refolding methods and different operating conditions in two gradients gel filtration process were investigated with regard to increasing the NTA protein activity recovery and inhibition of aggregation. The refolding of denatured NTA protein showed this method could significantly increase the activity recovery of protein at high protein concentration. The activity recovery of 37% was obtained from the initial NTA protein concentration up to 20 mg/ml. The conclusions presented in this study could also be applied to the refolding of lysozyme. |
| Databáze: | OpenAIRE |
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