Purification of recombinant human interferon gamma from fermentation broth using reverse micellar extraction: A process optimization study

ABSTRACT

In the present investigation, we have shown a single step purification of IFN-γ from the fermentation broth of recombinant Kluyveromyces lactis (K. lactis) strain, using cationic surfactant based reverse micellar extraction (RME) system. The forward and back extraction processes were optimized in order to improve the overall extraction process. 78%, 93% and 98% forward extraction efficiencies (FEE) were obtained after the process by using aqueous phase pH 12, 150 mM CTAB and 0.2 M NaCl respectively. Afterwards, back extraction efficiency (BEE) was maximized by the optimization of stripping phase pH, isopropyl alcohol (IPA) and potassium chloride (KCl) concentration by employing Taguchi’s method of orthogonal array. Stripping phase pH 7, 15% IPA & 0.8 M KCl were found to be the best possible parameters producing 83% BEE. The usage of IPA has proven to reduce the effect of micellar-micellar based interaction, which was reflected as an improvement in BEE. This study demonstrates that the reverse micellar system is a highly promising and efficient tool for the direct extraction of recombinant proteins from fermentation broth. Moreover, this system holds a high potential to be an integral part of the downstream process used in biopharmaceutical industry.

KEYWORDS:

• Interferon gamma
• reverse micellar extraction
• Taguchi’s method
• Kluyveromyces lactis

Acknowledgements

Rajat and Ashish would like to acknowledge the financial support extended by Institutional Doctoral fellowship by IIT Guwahati.

Declaration of interest

Authors declare that they have no conflict of interests.

Funding

This work was supported by a research grant from Indian council for medical research (Project no.: 2012-14790).

Additional information

Funding

This work was supported by a research grant from Indian council for medical research (Project no.: 2012-14790).