alexa Expression and Purification of SAK-fused Human Interferon Alpha in Escherichia coli
ISSN: 1948-5948

Journal of Microbial & Biochemical Technology
Open Access

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Research Article

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

Shardul Salunkhe, Bhaskarjyoti Prasad, Ketaki Sabnis-Prasad, Anjali Apte-Deshpande, Sriram Padmanabhan*

Lupin Limited (Biotechnology division), Gat no. 1156, Village Ghotawade, Taluka Mulshi, District Pune, India 411042

*Corresponding Author:
Dr. Sriram Padmanabhan
Lupin Limited (Biotechnology division)
Gat no. 1156, Village Ghotawade, Taluka Mulshi
District Pune, India 411042
Tel: +91-20-66549801
Fax: +91-20-66549807
E-mail: [email protected] lupinpharma.com

Received Date: December 04, 2009; Accepted Date: December 14, 2009; Published Date: December 15, 2009

Citation: Salunkhe SS, Prasad B, Sabnis-Prasad K, Apte-Deshpande A, Padmanabhan S (2009) Expression and Purification of SAK-fused Human Interferon Alpha in Escherichia coli. J Microbial Biochem Technol 1:005-010. doi: 10.4172/1948-5948.1000002

Copyright: © 2009 Salunkhe SS, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 

Abstract

A method for improved refolding and purification of E. coli derived human Interferon -? (rhIFN ?2b) from inclusion bodies as a Staphylokinase (SAK) fusion protein is described. Such a fusion protein did not require the supplementation of rare codons for expression and was found to be stable at 37?C. The optimal conditions of refolding involved the use of a mild denaturating agent without the need for any other agents to prevent aggregation. The SAKrhIFN ?2b fusion protein was successfully purified using two steps of purification and was cleaved using enterokinase 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 cleavage. The expression levels of SAK-IFN were found to be two folds higher than that observed with untagged IFN under similar experimental conditions.

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