Systematic optimization of active protein expression using GFP as a folding reporter

Protein Expression and Purification
Kentaro OmoyaNaomi Kondo

Abstract

Many recombinant proteins have been used as drugs; however, human proteins expressed using heterologous hosts are often insoluble. To obtain correctly folded active proteins, many optimizations of expression have been attempted but usually are found to be applicable only for specific targets. Interleukin-18 (IL-18) has a key role in many severe disorders including autoimmune diseases, and therapeutic approaches using IL-18 have been reported. However, production of IL-18 in Escherichia coli resulted in extensive inclusion body formation and previous conventional screenings of expression conditions could obtain only a condition with a low yield. To address the problem, we applied a folding reporter system using green fluorescent protein (GFP) for screening of the expression conditions for hIL-18. The established system efficiently screened many conditions, and optimized conditions for the expression of hIL-18 significantly enhanced the final yield of the active protein. Systematic screening using a GFP reporter system could be applied for the production of other proteins and in other organisms.

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Citations

Jul 15, 2009·Journal of Industrial Microbiology & Biotechnology·Nandadeva YakandawalaSrinivasa Madhyastha
Nov 5, 2013·Protein Expression and Purification·Mei M Whittaker, James W Whittaker
May 17, 2011·Microbial Cell Factories·Tomohiro MakinoGeorge Georgiou
Dec 11, 2007·Biochemical and Biophysical Research Communications·P DidierL Guidoni
Feb 3, 2007·Journal of Peptide Science : an Official Publication of the European Peptide Society·Sunmi KangSunghyouk Park
Sep 20, 2005·Analytical Biochemistry·Renaud VincentelliChristophe Bignon
Sep 1, 2005·Biochemical and Biophysical Research Communications·Henri R NordlundMarkku S Kulomaa
Nov 1, 2006·Allergology International : Official Journal of the Japanese Society of Allergology·Eiji MatsukumaNaomi Kondo
Jan 1, 2012·Computational and Structural Biotechnology Journal·Thorben Dammeyer, Philip Tinnefeld
Aug 25, 2009·Analytical Biochemistry·A Nicole EdwardsJennifer L Morrell-Falvey
Apr 6, 2016·Journal of Biomolecular Structure & Dynamics·Kseniya F GlukhovaBogdan S Melnik

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