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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Zhang, Yan Saremirad, Pegah Wood, Jeffery A. Ray, Ajay K. |
| Description | Country affiliation: Canada Author Affiliation: Saremirad P ( Department of Chemical and Biochemical Engineering, University of Western Ontario, London, Canada.); Wood JA ( Department of Chemical and Biochemical Engineering, University of Western Ontario, London, Canada.); Zhang Y ( Faculty of Engineering & Applied Science, Memorial University of Newfoundland, St John's, Canada.); Ray AK ( Department of Chemical and Biochemical Engineering, University of Western Ontario, London, Canada. Electronic address: aray@eng.uwo.ca.) |
| Abstract | Size exclusion chromatography has been demonstrated as an effective method for refolding a variety of proteins. However, to date process development mainly relies on laboratory experimentation of individual factors. A robust model is essential for high-throughput process screening and optimization of systems to provide higher productivity and refolding yield. In this work, a detailed kinetic scheme of oxidative refolding of a model protein (lysozyme) has been investigated to predict the refolding results in SEC. Non-reactive native, quenched and equilibrium studies were conducted to obtain the model parameters for the species formed during refolding of denatured/reduced lysozyme. The model was tested in various operating conditions, such as: protein loading concentration, injection volume, flow rate and composition of refolding buffer with and without the use of l-arginine additive. An apparent two-state mechanism was found adequate to describe refolding of lysozyme on SEC for the operating condition tested in this work. Furthermore, using low concentration of l-arginine combined with urea as common aggregation suppressor additives showed insignificant change in kinetics of refolding of lysozyme on SEC. However, addition of l-arginine changed mass transfer properties of some of the species formed in refolding reaction which was considered in the model to accurately predict the result of refolding on SEC. |
| ISSN | 00219673 |
| Volume Number | 1370 |
| e-ISSN | 18733778 |
| Journal | Journal of Chromatography A |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2014-11-28 |
| Publisher Place | Netherlands |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Chromatography, Gel Methods Muramidase Analysis Protein Refolding Kinetics Models, Biological Chemistry Oxidation-reduction Journal Article Discipline Analytical Chemistry |
| Content Type | Text |
| Resource Type | Article |
| Subject | Organic Chemistry Medicine Analytical Chemistry Biochemistry |
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