The BioLab has chosen to highlight the current student projects in the lab, and the now-ready AF4 system.
Contents:
Equipment Update
The AF4 system is now available
The
Wyatt Eclipse AF4 Field-Flow Fractionation (FFF) System discussed in the
last issue is now setup and ready for use by the user programme. As a reminder this equipment can be used to seperate baculovirus particles and lopid nanoparticles and distinguish empty from full. This is similar to a HPLC, but allows for more freedom, as it seperates by mass rather than particle size. This system will soon be commissioned on the SANS2D beamline with the help of Najet's industrial placement student, due to start in July 2024.
We also have a fraction collector for the AF4, so we can now collect and measure different particle size materials rather than just the end product.
Studentships
There are currently 2 student-based biodeuteration projects running in the Biolabs currently. The first is a sandwich placement student (Luke
Broughan, finishing in September). The second James Goon who will finish his year placement in August.
Luke
Broughan, sandwich placement student (Joint project with Peixun)
Luke's project involves sample synthesis, growing bacteria in deuterated media. Bacteria can generally only handle a small amount of deuterated media, so he has grown it in stages, increasing the deuterated percentage of the media and selecting from the most well-adapted bacteria to grow the next stage. This results in bacteria adapted to growing in a highly deuterated media, which can then be processed further to obtain the deuterated bacterial membranes.
The ultimate aim of this project is to obtain deuterated bacterial lipopolysaccharides (LPS), which will be used on beamlines as a model for the bacterial membrane, for example Luke Cliffton's reflectometry measurements to probe antibacterial drugs using a membrane model utilise purified LPS in building the surfaces.The value of using this biochemical purification method of outer bacterial membrane is that it doesn't require a large quantity of harsh chemicals such as phenol, chlorofom etc. The resulting LPS will be significantly cleaner when compared to the current chemical extraction methodologies.
More details about Luke's placement are available here. A copy of Luke's presentation is available here.
James Goon, sandwich placement student (Joint project with Najet)
James' project is similarly involved in the synthesis of biodeuterated proteins, looking at both baculoviral expression system using insect cells. These insect cells are closer to animal cells in how the proteins are modified and folded, which makes them valuable for more accurate models. However, D2O is toxic for them, so the only way to deuterate them is to use a deuterated carbon source. Deuterated amino acids are expensive so we are looking to use processed deuterated algae extract, produced at ESS by Zoe Fisher. Recombinant baculovirus protein by comparison is closer in resemblance to mammalian protein than bacterial recominant protein and is better for working at larger scales, allowing protein production at the scale needed for neutron beamline measurements. Once succesful, we will be using deuterated viruses as well as deuterated recombinant proteins produced in baculovirus on the SANS2D beamline, studying protperties of protein complexes, as well as structure of viruses.
Summer Placement Student (Sian Pritchard)
A summer placement student (Sian) has been selected from the many applicants. The project will run for 12 weeks, and will involve biodeuterated algae: these algae can be 'juiced' to provide a deuterated carbon source for insect cell/baculovirus sysyem, effectively acting as deuterated feed. We will also use the deuterated algae extract to produce recombinant proteins for users and our collaborators at CLF who will use deuterated fluorescent proteins to study proton tunneling quantum effects. Sian is enrolled on a biochemistry masters at University of Southampton.
General Upates:
SAS EMBL course
Carrying on with the theme of support for the SANS beamlines, the Biolab manager has recently attended a course focused on small angle scattering techniques and as such can now provide additional support to proper sample preparation for measurements of biomolecules for small angle scattering users. Additionally, this allowed the Biolab manager to form working relationships with colleagues at DESY in Hamburg.
AILM2024
The Biolab manager has also attended AILM2024 (link: https://www.ailm2024.org/ ) and presented a poster (attached), which featured the work conducted by Luke and James during their projects.
Presented poster at AILM2024.
