Background

Many proteins and enzymes of biotechnological importance contain structure stabilizing disulphide bonds. Currently these proteins are difficult for the biotech industry to produce on a large scale. Often refolding or production in mammalian cells is needed.

Currently available bacterial strains whose cytoplasm is more oxidizing and thus more favorable for disulphide bond formation are inefficient systems as there are no active mechanisms for disulphide bond formation. Hence from these strains proteins products are unable to attain their native conformation and they form insoluble aggregates.

Technology Overview

Researchers at Oulu have invented the CyDisCo, Plasmid based system with 1 to 3 protein folding catalysts.

Developmental Status

TRL 5‑6 (Technology validated/demonstrated in relevant environment)Successfully tested with over 400 different proteins including full length antibodies and Fab-fragments internationally by academia and industry.

References

• Hatahet F, Nguyen VD, Salo KE, Ruddock LW Microb Cell Fact 2010: 9-67.
• Nguyen VD, Hatahet F, Salo KE, Enlund E, Zhang C, Ruddock LW Microb Cell Fact 2011:10:1.

Benefits

• Production of active recombinant proteins having disulphide bonds in the cytoplasm of bacteria
• No need for refolding
• Works with any E. coli strains and in any media
• Allows large scale production and processing
• Up to 800 fold increase in yield of active protein
  • Up to 1 g / L in fed-batch cultures
  • Up to 150 mg / L full length antibodies in shake flasks

Applications

• Production of proteins (e.g. enzymes, antibodies, Fabs, ScFvs) containing disulphide bonds (up to 11) used as drugs, enzymes, components of washing powder and diagnostics, etc

Opportunity

• Switch away from mammalian cells and inefficient alternative production systems
• Free non‑exclusive trial period of max. 6 months: Test with your proteins to see how much more active protein you can get.
  • Excluding pharma and veterinary use (currently reserved for exclusive evaluation)
• Available for licensing (different applications and different optional terms)
• Collaborative R&D projects (new proteins)