DPhil in Clinical Medicine: New approaches to vaccine development against Epstein Barr Virus and Rabies
Lead Research Organisation:
University of Oxford
Department Name: Clinical Medicine
Abstract
The role of the viral Membrane Fusion Protein (MFP) is to force the lipid bilayer of the virus membrane into close enough proximity to that of the host cell that they merge, facilitating entry of the virus. This fusion relies on significant conformational changes in the MFP, from a pre-fusion to a post-fusion conformation, to overcome the energetic penalty associated with the similarly-charged membranes being brought together. It might therefore be possible to prevent viral entry into the cell by inhibiting the action of the MFPs, either by blocking binding to its relevant host cell receptor, or trapping it in the pre-fusion conformation. This project aims to elucidate how the pre-fusion and post-fusion structures of two MFPs, Rabies Virus Glycoprotein and Epstein Barr Virus glycoprotein B, might be manipulated for use in vaccine development against these two viruses. By promoting the generation of antibodies that recognise these MFPs, such vaccines might elicit protection against cell entry of the virus and therefore against disease.
People |
ORCID iD |
| Edward English (Student) |
Publications
Ng WM
(2022)
Structure of trimeric pre-fusion rabies virus glycoprotein in complex with two protective antibodies.
in Cell host & microbe
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| MR/N013468/1 | 30/09/2016 | 29/09/2025 | |||
| 2436253 | Studentship | MR/N013468/1 | 30/09/2020 | 29/09/2024 | Edward English |
| Description | Monoclonal Antibody isolation against viral targets using a Berkeley Lights Beacon |
| Organisation | University of Edinburgh |
| Department | Edinburgh Genome Foundry |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Our collaboration with the Edinburgh Genome Foundry seeks to isolate antibodies against viral protein targets. Toward this, we are using their state-of-the-art screening technology, the Beacon (Berkeley Lights, California), which is able to screen individual immune cells for 'hits' that make antibodies against the desired target. My contribution to this collaboration is to develop immunogens that will stimulate a response in mice, extract and process the cell samples from these immunised mice so they can be run on the machine, perform optimisation experiments testing the best way to handle these cells, including how to enrich for the specific cell types of interest (antibody-secreting cells), experimental design when using the Beacon and optimisation of the downstream cell processing pipeline. Biological characterisation of any antibodies we isolate from this experimental work will also be done by me. This work, including the planning and troubleshooting, is overseen by my PhD supervisor in Oxford. |
| Collaborator Contribution | The Edinburgh Genome Foundry (EGF) have kindly provided us with the consumables and reagents we require to perform experiments on the Beacon (which they also own). The Beacon itself is a complex technology that requires specialist training to operate - hence, all of the machine-based steps in the antibody discovery pipeline are conducted by the facility specialist there. The facility manager provides supervision and support for the experimental work we conduct there. We have previoulsy made brief use of other core facilities at the University of Edinburgh to support the Beacon work, including the Flow Cytometry Facility and the Roslin Institute Animal House, though these aspects of the experimental pipeline now have simpler workarounds that I can perform myself. |
| Impact | So far, we have spent the first year of our collaboration working on setting up and streamlining the experimental pipeline. This has predominantly yielded useful information on how to maximise the number of 'hits' per run, using the minimum amount of animal cell samples as possible. Now that the pipeline is well set up, we are starting the process of isolating antibody sequences which will be re-expressed for downstream characterisation, though this is in its early stages. |
| Start Year | 2022 |