Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10236017 | Process Biochemistry | 2011 | 5 Pages |
Abstract
Expression of recombinant proteins in E. coli often results in formation of inclusion bodies and need to undergo renaturation. The effect of parallel feeding of oxidizing agent and denatured protein was investigated to control side oxidation and refolded yield of interferon beta-1b. Different molar ratios of iodosobenzoic acid (IBA) and interferon beta-1b (IFNβ-1b) were applied using a fed-batch process. The Reverse-Phase HPLC method was utilized to differentiate the native IFNβ-1b from the non-native oxidized and related proteins formed during the refolding process. The concentrations of IBA influenced native to non-native protein ratio hence affecting the protein quality and yield. The oxidized and related proteins were affected reversely. High concentrations of IBA increased the formation of oxidized protein while at low concentrations there was a slight increase in related proteins. The amount of 40 μg mlâ1 of IFNβ-1b and 8 μM of oxidizing agent was found to favor the refolding process.
Related Topics
Physical Sciences and Engineering
Chemical Engineering
Bioengineering
Authors
Ahmad Fazeli, Seyed Abbas Shojaosadati, Mohammad Reza Fazeli, Hooshmand Ilka,