Project No. 2425
Prof Michelle D. Garrett – University of Kent
Dr Tim Fenton – University of Southampton
Prof Dan Mulvihill – University of Kent
Production and delivery of biologically active recombinant proteins into target cells is still an important ongoing challenge in biotechnology.
Production and delivery of biologically active recombinant proteins into target cells is still an important ongoing challenge in biotechnology. We have developed a transformative technology to solve this problem, which utilises an innovative recombinant E.coli-based expression system to induce packaging of a range of recombinant proteins in membrane bound vesicles. This enables production of otherwise challenging insoluble, toxic, disulphide bond containing recombinant proteins in bacteria. Preliminary studies with these vesicles show that they can fuse with human cells in culture, thus delivering their cargo into the cell. The aim now is to modify the surface of these vesicles so that we can selectively target them to particular cell types.
The proof of concept will be to express the PD-1 protein, which binds to PD-L1 expressed on specific types of human cells, on these vesicles (Kent) with a recombinant GFP protein as the vesicle cargo. These vesicles will then be isolated and incubated with different human cell types that do/do not express PD-L1 to evaluate cellular selectivity and impact (Kent), and analyse gene expression changes by RNA sequencing (Southampton).
Areas of potential impact:
The potential impact of this project is that it will deliver proof-of-concept that a biologically active protein can be recombinantly expressed in E.coli generated vesicles and selectively targeted to enter a specific cell type. This is a transformative technology that has the potential to be applied to help address needs in both basic biological science research into cell function, and the biotechnology industry. In this project the student will gain experience in a wide range of cell biological, molecular, biophysical and biotechnological techniques, working with researchers and academics with a wealth of experience in these areas.
An interest in and/or experience of protein biotechnology in the laboratory setting and human cell culture.