Expression and Purification of SAK-fused Human Interferon Alpha in Escherichia coli

Abstract

See, stats, and : https :/ / www. researchgate . net / publication / 45459467 Expression - fused Human Article DOI : 10 . 4172 / 1948 - 5948 . 1000002 : DOAJ CITATIONS 4 READS 101 5 , including : Shardul Lupin , Inc . 6 SEE Bhaskarjyoti Lupin , Inc . 5 SEE Anjali - Deshpande Lupin , Inc . 22 SEE Sriram CanCyte . Ltd 47 SEE All . The . All - text and , letting . Abstract A method for improved refolding and purification of E . coli derived human Interferon - α (rhIFN α2b) from inclu - sion bodies as a Staphylokinase (SAK) fusion protein is described . Such a fusion protein did not require the supple - mentation of rare codons for expression and was found to be stable at 37 o C . The optimal conditions of refolding in - volved the use of a mild denaturating agent without the need for any other agents to prevent aggregation . The SAK - rhIFN α2b fusion protein was successfully purified using two steps of purification and was cleaved using enteroki - nase into two fragments namely SAK and IFN . Both the proteins were found to be biologically active showing proper folding of both the fusion partners . The cleaved IFN showed similar retention time on RP - HPLC as the bacterial derived untagged purified IFN as well as similar molecular weight on Agilent 2100 Bioanalyzer indicating the right processing of the IFN after enterokinase cleav - age . The expression levels of SAK - IFN were found to be two folds higher than that observed with untagged IFN under similar experimental conditions .