Inhibition of aggregation by media selection, sample loading and elution in size exclusion chromatographic refolding of denatured bovine carbonic anhydrase B.
The effects of gel media, sample application and elution flowrate on the activity recovery and aggregation in refolding of bovine carbonic anhydrase B (CAB) by size exclusion chromatography (SEC) were investigated. Variation in aggregation was demonstrated visibly by the comparison of refolding profiles under different operating conditions. Some principles with regard to practical application were proposed. Meanwhile, the analysis of relationship between peak resolution and activity recovery provided evidences for the mechanism of size exclusion chromatography protein refolding.
Gu
Zhenyu
Z
Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, PR China.
Zhu
Xiaonan
X
Ni
Shaowei
S
Zhou
Haimeng
H
Su
Zhiguo
Z
D000818:Animals; D024401:Carbonic Anhydrase I / Q000737:chemistry* / Q000302:isolation & purification*; D002417:Cattle; D002850:Chromatography, Gel / Q000379:methods*; D004789:Enzyme Activation; D005782:Gels / Q000737:chemistry*; D046911:Macromolecular Substances; D044085:Microfluidics / Q000379:methods*; D011485:Protein Binding; D011489:Protein Denaturation; D017510:Protein Folding
D005782:Gels; D046911:Macromolecular Substances; D024401:Carbonic Anhydrase I