Annaig Rozo

During my Bachelor degree I spent my final year as an Erasmus student at Aston University. I was awarded a Master's degree in Cancer Biology from the University of Lyon in 2019 during which I undertook two research internships. The first one was at the University of Tokyo where I was working on the role of inflammatory monocytes in the extravasation of breast cancer cell. The second internship was at the Aston University where I focused on the characterization of extracellular vesicles and their role in recruiting macrophages during inflammation.

Project: Characterisation of extracellular vesicles using microfluidic approaches

Extracellular vesicles (EV) are released from cells and act as a conduit for transfer of signals between them. However, relatively little is known about how EV act or the key modulators that underpin their function. Due to their small nature, traditional purification methods generally involve high-force regimes e.g. ultracentrifugation. Coupled to this, populations of EV are highly heterogeneous both in terms of size and composition. A key area for understanding is the varying functional effects of these different EV and the molecular basis underpinning their activity.

This project will utilise bespoke proprietary microfluidic chip technology to separate EV from cells and to sub-fractionate them by size under low-force regimes. These chips have been developed by the industrial partner, uFraction8, and have been demonstrated to be capable of separating biological particles on the nanoscale. Once separated, EV population will be subjected to rigorous biochemical (e.g. lipid and protein composition), biophysical (e.g. size, surface charge, membrane fluidity) and functional assays (e.g. control of inflammation). Taken together, these analyses will reveal the underlying properties of different EV populations that are responsible for their activities. This will pave the way to a better understanding of both normal and disease-state EV activity as well as the development of novel EV-based therapeutics.

Athanasios Kesidis

My studies began in Greece where I completed an undergraduate degree in Molecular Biology and Biotechnology Sciences at the University of Crete. During my research traineeships in Greece and England, I appreciated the significance of membrane proteins and their role in an incredible array of functions in the human body. For my undergraduate thesis, I investigated the role of cysteine 184 in the structure, function and the expression of the type 2 receptor (CRF2) of corticotropin-releasing factor. My future career goal is to become a world-leading expert in the pharmacology of membrane proteins and specifically G protein-coupled receptors. Furthermore, I want to be part of cutting-edge projects which will provide the world with all the information that is needed to find cures and treatments for major diseases which plague our society.

Project: Optimisation of membrane protein expression and purification

Peak Proteins Ltd. is involved in the production and supply of bespoke high-quality protein reagents to clients and the delivery of macromolecular structure solutions of client targets, largely to aid pharmaceutical and drug discovery research.

The student will be helping to expand Peak Proteins capabilities to handle membrane protein targets. They will be involved in the development of a rapid, simple and generic approach to membrane protein stability assessment using fluorescent protein tags. The project will include the expression of a number of therapeutically relevant membrane protein targets from different classes, with the initial focus on a serotonin receptor (GPCR) and an iron-sulphur cluster transporter (ABC transporter). Appropriate expression systems will be used, including E. coli, insect cell and HEK systems. New approaches will be benchmarked against traditional methods for assessing membrane protein quality, such as radioligand binding assays, protein activity assays and transport assays.

The student will gain a broad overview of protein production pipelines, from gene construct design, through expression, to protein purification methods. Furthermore, the student will gain specific skills such as tissue culture, membrane protein solubilisation, and biophysical assays for membrane protein assessment. Finally, the student will gain a unique perspective on a career in commercial research from a contract research organisation perspective, including risk management and business development skills.

Alexis Lode

I completed a BSc in Biology in Rennes, France, during which I was able to follow my final year in Erasmus exchange in Aston University in Birmingham. Then, I continued with a MSc in Conception of Therapeutic and Diagnostic Tools in Bordeaux, France. I am very interested in drug development against multiple targets, and I particularly liked my time in Birmingham, which is why I decided to join the AMPL team from October 2019 to do a PhD based on the use of membrane proteins as antifungal drug targets. This project aims to use innovative purification techniques (SMALPs) in order to purify membrane proteins that will have a certain affinity for potential antifungal drugs.

Project: Membrane proteins as antifungal drug targets

An estimated 1.5 million people die from invasive fungal infections each year. With limited treatment options and increasing resistance to available therapies, there is an urgent need for new antifungal drugs acting via novel mechanisms of action. To address this issue, this project will investigate membrane proteins as antifungal drug targets. F2G Ltd have antifungal programmes at different stages that target membrane proteins. The phase 2 antifungal compound olorofim acts by inhibiting the mitochondrial membrane protein dihydroorotate dehydrogenase. Additionally, a multi-spanning membrane protein has been identified as the target of a preclinical series of antifungals. Further potential membrane protein targets will also be assessed in this project. Traditionally, membrane proteins have been difficult to work with so that preparation of recombinant versions of potential drug targets for inhibitor screening has not always been possible. However, the use of innovative purification techniques including SMA lipid particles (SMALPs) has enabled difficult membrane proteins to be purified in their native form allowing their activity to be studied.

This project will apply cutting edge membrane protein technology to the exploration of antifungal drug targets, with the aim of supporting existing drug programmes and identifying potential novel targets.

Peer Depping

Peer studied chemistry at the university of Bonn in Germany where he received his bachelor’s degree in 2016 and his master’s degree in 2018. During his studies he worked on the crystallization of the trityl labelled cytochrome P450cam and the functional and structural analysis of the tripartite ATP-independent periplasmic (TRAP) transporter SiaPQM using nanodiscs and liposomes as membrane mimetic systems. His topic in the MemTrain programme is the development of antibody-based drugs targeting ion channels.

Project: Development of antibody-based drugs targeting ion channels

Ion channels represent the second largest family of human cell surface proteins and are implicated in a number of diseases including cancer, autoimmunity and chronic pain. To date, most of ion channel drug discovery focused on developing small molecules and peptides as therapeutics. These drug modalities often suffer from poor selectivity for the target ion channel leading to severe side effects and poor efficacy in patients. An attractive alternative is the use of antibody-based therapeutics which can demonstrate superior affinity, selectivity and in vivo half-life. However, the efforts to generate monoclonal antibodies against ion channels are hampered by the difficulties in expression and purification of ion channels in a format suitable for antibody isolation and screening.

Iontas is an innovative antibody-drug discovery company developing a novel platform technology (KnotBodyTM) targeting ion channels using antibody-like molecules. As part of the development of this technology, purified ion channels are extremely desirable. As such, this project will focus on expression, purification and functional analysis of ion channels, using cutting-edge polymer-based technology developed at Aston University. Following this, there will be the opportunity to undertake a substantial placement at Iontas to generate and validate antibodies against the ion channel(s) of interest using phage and/or mammalian display technology. Ultimately, this will aid the development of novel therapeutics.

Afroditi Vaitsopoulou

I studied Biochemistry and Biotechnology in Greece where I also worked in the laboratory of bio-organic chemistry of my university. Then, I did an Erasmus + internship at Cancer Research Center of Marseille, in France. Moreover, I did my master in Marseille, on bioinformatics and structural biochemistry, where I did my internship in the laboratory of viral replicases. Always interested in different domains and multidisciplinary fields, I am happy to be in this team and to have the flexibility to move between the various fields. My project during this program is to find the best approach for stabilizing and characterizing structurally and functionally wild type membrane proteins. I am currently working on MRP4, a protein that is resistant to drugs including cancer chemotherapy, antivirals and antibiotics.

Project: Approaches for stabilising wild type membrane proteins for structural and functional characterisation

Membrane proteins are important drug discovery targets for a wide range of diseases. However elucidating the structure and function of native membrane proteins is notoriously challenging. Detergents have been used to solubilize membrane proteins from the lipid bilayer with varying degrees of success concerning protein stability. To try to improve stability many studies use mutagenesis approaches to further stabilise the protein, but this can often affect the structure and function of the protein. CALIXAR is a French SME which specialises in purification technologies for membrane proteins. They have developed a series of stabilising detergents and additives which have been shown to stabilise extracted membrane proteins, maintaining the native conformation in solution.

This collaborative project will include cryo-electron microscopy structural characterization of human transporter protein MRP4 (multidrug resistance protein 4/ABCC4), solubilised and purified using calixarene-based detergents and /or styrene maleic acid (SMA) polymers. In addition novel reconstitution approaches will be investigated to monitor MRP4 function. Finally the project will involve testing of novel polymers and additives developed by Calixar/Aston to evaluate their potential in terms of solubilisation efficiency, stabilization, function and compatibility with downstream techniques.