The research – which will be funded by a £217k ($245k) grant from Innovate UK and the BBSRC – will see scientists from Cobra work with counterparts at the University of Manchester to optimise E.coli for industrial applications.
The main focus will be using the University’s RiboTite translation inhibition technology to make sure E.coli cells used for protein production only make as much as they can secrete according to Cobra chief scientific officer (CSO), Daniel Smith.
“A lot of protein can be made in a relatively short time frame in E.coli, which can result in the formation of inclusion bodies in the cytoplasm of the host cells” Smith told Biopharma-reporter.com, adding that removing these protein aggregates is costly and time consuming.
Neil Dixon from the University of Manchester agreed, telling us overloading of the secretion system can be related to "the upstream ancillary proteins or indeed the pore responsible for the actual translocation."
"The RiboTite expression system is composed of an engineered E. coli strain and series of expression plasmids. The unique feature is that it uses mRNA devices, or orthogonal riboswitches, that operate at the level of translation initiation. This feature allows tight control of basal expression in the absence of induction and tuneable control of expression across a broad dynamic range" Dixon added.
Right place, right time
The other problem is that proteins only adopt the correct structure in certain parts of the cell.
Smith explained that the disulphide bonds that give many therapeutic molecules their shape only form in proteins secreted into the periplasm and not in those trapped in the cytoplasm.
“We are looking to develop a system that allows for fine control of expression of a target protein coupled to periplasm targeting, in order to overcome the burst of expression and also to allow for the correct formation of disulphide bonds.
“An additional benefit is that the protein content of the periplasm is less enriched in contaminating proteins then the cytoplasm which can also help with the purification steps during the manufacture” Smith said.
To me and my bank manager, £217k is a lot of money. But to a contract manufacturing organisation like Cobra it is a relatively small sum, however, the project is still a good use of the firm’s time according to CEO Peter Coleman.
He told us Cobra “likes to work in collaboration with other scientists to achieve its R&D goals. Grants like this from the BBSRC and Innovate UK allow scientists from both industry and academia to develop products and technology together.
Colman added that such projects allow “Cobra to evaluate technology solutions that exist outside of the company that could potentially enhance our Microbial Protein Toolbox service offering.”