The BioLab has continued to acquire new capabilities, projects and students.
Contents:
New Equipment:
Shimadzu High Pressure Liquid Chromatography (HPLC)
The Shimadzu HPLC is available for use. While this is used on
the
SANS2D beamline, the offline optimisation work is conducted in the Biolabs, and it is considered part of the Biolab equipment, allbeit optimised for beamline measurements.
This equipment consists of an autosampler, which feeds into a sample seperation column. These seperated samples are measured with a UV detector to correlate the fraction being measured with the beamline measurement; the sample is then measured directly on the beamline. Due to the autosampler tray, multiple samples can be measured in sequence, making it much more efficient to run multiple measurements.
HPLC in place on SANS2D Beamline.
Tom Cooper, Najet's industrial placement student for 2022-2023, has been working extensively with the Shimadzu HPLC, and is the current local expert in its operation. Tom will also be conducting some initial experiments on the Wyatt Eclipse system mentioned below.
Wyatt Eclipse AF4 Field-Flow Fractionation (FFF) System
The Wyatt Eclipse AF4 Field-Flow Fractionation (FFF) System has been delivered and installed in the Target Station 2 Biolab. It is ready for commissioning. It is expected that it will be used in future projects by the new industrial placement students and the new PhD student.
Wyatt Eclipse AF4 Field-Flow Fractionation (FFF) System (left) and Eclipse unit specifically (right).
The FFF system utilises a set of orthogonal flows to isolate media based on mass; lighter particles are subjeced to different flow rates compared with heavier particles, resulting in their seperation. As such, the new FFF System will allow for far more freedom in particle size when compared with the HPLC which seperates by particle size; it is a complementary process to Size Exclusion Chromotography (SEC).
Wyatt Eclipse AF4 Field-Flow Fractionation (FFF) System in operation.
The FFF System will also be able to be coupled to SANS2D, allowing for in-situ measurements on phase-seperated media, but can currently be used offline.
Studentships
2023/2024 Industrial Placement students
The two industrial placement students in the BioLabs mentioned in the previous newsletter started in August and September. The BioLab would like to extend a warm welcome to Luke Broughan and James Goon. Luke is focused on the biodeuteration of bacterial membranes - a joint project with Peixan, and line managed by Ludmila. James is focused on the protein expression and biodeuteration in insect baculoviral systems – a joint lab project with Najet, who is also their line manager.
PhD Studentship (2023-2027) between ISIS, Oxford Brookes University and Oxford Expression Techologies
"Investigating the structure of extracellular vesicles and budded virus in baculovirus replication"
The BioLabs jointly with Oxford Brookes University and Oxford Expression Technologies have secured funding for a PhD studentship starting in 2023; Part of this project will be based in the ISIS BioLabs, and part based at Oxford Brookes University, with some time at OET.
The PhD is also based on
baculovirus expression, much like the BBSRC funding, specifically the project outline involves using Small Angle Neutron Scattering (SANS) and cryogenic electron microscopy to investigate virus titre levels with respect to virus structure to optimise the useful protein production.
It will involve extracellular vesicles, specifically exosomes, particles used by cells to communicate between cells over medium and long distances.
The overall project supervisor is Professor Linda A King from Oxford Brookes University, with Dr Najet Mahmoudi from ISIS and Dr Adam Chambers also acting as supervisors.
General Upates:
BBSRC Funding
Najet along with Oxford Expression Technologies (OET) has been awarded a BBSRC facility access fund for bioscience partnerships based on the "Effect of recombinant protein expression on baculovirus budded virs structure and infectivity" for 2023. The application for this funding was based on the work of Leo P. Graves, Louise C. Hughes, Sarah L. Irons, Robert D. Possee, Linda A. King in their paper "In cultured cells the baculovirus P10 protein forms two independent intracellular structures that play separate roles in occlusion body maturation and their release by nuclear disintegrationior" (https://doi.org/10.1371/journal.ppat.1007827.g001). The experiment based on the facility access fun is planned to go ahead on SANS2D in July.
