Step change of mobile phase flow rates to enhance protein folding in size exclusion chromatography
Resource
Biochemical Engineering Journal 29(2006), 2–11
Journal
Biochemical Engineering Journal 29(2006), 2–11
Pages
-
Date Issued
2006
Date
2006
Author(s)
DOI
246246/2006111501254434
Abstract
Minimal aggregation formation between injector and column inlet and enough residence time inside the column were identified to be the
key for proper proteins refolding by size exclusion chromatography (SEC). Therefore, a step change of mobile phase flow rate strategy was
developed. That is, during the injected denatured protein (sample) traveling from injector to column inlet, a higher rate of mobile phase should
be applied to reduce aggregation. Then, a lower flow rate should be used to allow enough time for protein to refold inside the column. The
refolding of denatured lysozyme in SEC by this method was shown to reduce the aggregation and thus resulting in good refolding yields,
though not completely. Combining this method together with our previously proposed chaperon solvent plug strategy could obtain complete
recovery of denatured protein, both mass and activity recoveries.
Subjects
Step change of mobile phase flow rates
Chaperon solvent plug
Size exclusion chromatography
Protein refolding
Lysozyme
Publisher
Taipei:National Taiwan University Dept Chem Engn
Type
journal article
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