title: Improving the refolding of NTA protein by urea gradient and arginine gradient size-exclusion chromatography.
abstract: 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.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Fan
authors.forename: Diansheng
authors.initials: D
authors.lastname: Xu
authors.forename: Bing
authors.initials: B
authors.lastname: Lu
authors.forename: Jie
authors.initials: J
authors.lastname: Xia
authors.forename: Dongzhi
authors.initials: D
authors.lastname: Wei
mesh_terms: D001120:Arginine / Q000494:pharmacology*; D002850:Chromatography, Gel / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005343:Fibrinolytic Agents / Q000737:chemistry* / Q000378:metabolism; D009113:Muramidase / Q000378:metabolism; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding; D020673:Protein Renaturation; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014508:Urea / Q000494:pharmacology*
chemical_list: D005343:Fibrinolytic Agents; D011994:Recombinant Proteins; D014508:Urea; D001120:Arginine; D009113:Muramidase