Researchers at VIB are cloning an IL15 gene into a range of VIB expression vectors but were producing insoluble proteins and consequently very low protein yields until they used this refolding system
Novexin, a company specialising in protein processing technology, reports that Belgium-based VIB (Flanders Institute of Biotechnology), one of Europe's leading biotechnology institutes, is successfully using Novexin's protein refolding system to generate excellent yields of a previously insoluble IL15 protein.
Researchers at VIB are cloning an IL15 gene into a range of VIB expression vectors known as FastScreen (Biotecnol SA) and are then expressing recombinant IL-15 in E coli strains including K12 and BL21.
They found both their wild type and mutant IL15 clones were producing insoluble proteins and consequently very low protein yields (less than one percent).
However, when they used the Novexin refolding technology in their purification process, protein yields increased by over 75 times.
Professor Nico Mertens, a project leader at VIB, explained: "We want to develop an inexpensive cloned pro-inflammatory cytokine, IL15.
"This protein is expressed in several inflammatory disorders, including rheumatoid arthritis and psoriasis and may therefore serve as a useful therapeutic target.
"We were having real problems obtaining any IL15 from our E coli preparations even using harsh denaturing conditions, which would normally solubilise most proteins.
"But when we used Novexin's refolding technology our yields increased dramatically, and because the reagents are non-detergent based the protein is active too.
"This means we can confidently use the Novexin refolding system in scale up, to develop a cost-effective manufacturing process for our IL15 protein".
Daniel Jones, CSO of Novexin, stated: "We are delighted to see a major biotechnology institute such as VIB obtaining such astonishing levels of yield increase.
"The VIB study is a good example of the results you can obtain with our system, which is now available as a simple to use kit and shows researchers working with seemingly insoluble therapeutic proteins how Novexin's refolding technology could significantly improve their protein-based research and manufacturing."