Inhibition of aggregation by media selection, sample loading and elution in size exclusion chromatographic refolding of denatured bovine carbonic anhydrase B
Department of Biochemistry and Molecular Pharmacology; Program in Molecular Medicine
Medical Subject Headings
Animals; Carbonic Anhydrase I; Cattle; Chromatography, Gel; Enzyme Activation; Gels; Macromolecular Substances; Microfluidics; *Protein Binding; Protein Denaturation; *Protein Folding
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.
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Citation: J Biochem Biophys Methods. 2003 Jun 30;56(1-3):165-75.
Gu, Zhenyu; Zhu, Xiaonan; Ni, Shaowei; Zhou, Haimeng; and Su, Zhiguo, "Inhibition of aggregation by media selection, sample loading and elution in size exclusion chromatographic refolding of denatured bovine carbonic anhydrase B" (2003). Biochemistry and Molecular Pharmacology Publications and Presentations. Paper 117.