Cytofold StructQuant^® Protein Folding Technology

Cytofold StructQuant^® protein folding is a proprietary technology of Paras Biopharmaceuticals, for the high-level expression of ho­mogeneously folded disulfide bond containing proteins in the cytoplasm of E. coli. The technology offers all of the cost benefits of expression in E.coli rather than in eukaryotic systems, while at the same time retaining the high yields obtained in cytoplasmic expression. The biologics produced are of the highest quality and have maximal biological activ­ity, unlike the activity levels obtained for many secreted eu­karyotic proteins when expressed in the cytoplasm without Cytofold StructQuant^® technology.

Native disulfide formation is a complex multi-factorial proc­ess and has not been possible in the cytoplasm of E. coli due to the presence of two reducing pathways. The synthesis of disulfide bond containing proteins is therefore either done in eukaryotic cells, in the periplasm of bacteria or by using in vitro oxidative refolding methods. All of these have prob­lems relating to cost and efficiency.

Technology allows:

• Low cost production of biologics that contain disulfide bonds.
• Large scale production of biologically active eukaryotic proteins in the cytoplasm of E. coli.
• Decreased production costs via high yield production in E. coli.
• Reduced process development time as in vitro refolding optimization is not required.

Disulfide bond formation is a post-translational modification that is required for the correct folding and function of around one-third of human proteins, including most secreted and outer-membrane proteins. Proteins that include disulfide bonds include insulin, antibody fragments, interleukins, interferons, erythropoietin and many others.

Native disulfide formation is a complex multi-factorial proc­ess and has not been possible in the cytoplasm of E. coli due to the presence of two reducing pathways. The synthesis of disulfide bond containing proteins is therefore either done in eukaryotic cells, in the periplasm of bacteria or by using in vitro oxidative refolding methods. All of these have prob­lems relating to cost and efficiency.

Unlike previous systems, this works in any E. coli strain and does not require the disruption of the reducing pathways naturally present. The system also works in many media compositions.

Paras Biopharmaceuticals Finland Oy uses Cytofold StructQuant^® for the development of its own products and in the development of production processes for other companies. Furthermore, Paras Biopharmaceuticals Finland Oy has in-house expertise in Cytofold StructQuant^® technology to ensure that the most appropriate variant is used to further cut the cost and time of development.