ADDRESS Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU UK
CONTACT e: t: 01865 613200
2021 Mark Howarth. All rights reserved.
The theme of our research is Innovating Protein Nanotechnologies for Immune Activation. We have a number of project areas running in the lab, ranging from fundamental analysis of protein interactions through to clinical application. Immuno-engineering and Global Health Developing an effective vaccine may be the most effective way to improve human health. We have established an approach to accelerate vaccine development, through our Plug-and-Protect platform. A limiting factor in vaccine generation is the difficulty of turning a promising target protein into the kind of assembly that would give long-lasting disease protection. We showed rapid and efficient decoration of virus-like particles, which elicited a strong immune response even with only a single injection. We have demonstrated potent immunization towards the global health challenge of malaria, working with collaborators at Oxford University’s Jenner Institute. This approach is now being used by many groups against cancer and various infectious diseases, e.g. HIV, influenza outbreak pathogens (including Covid-19), and veterinary diseases. The Plug-and-Protect platform has entered Phase I clinical trials for Covid-19, with trials for further diseases in preparation. Our lab’s current focus is the tailoring of antigens and protein cages, as well as establishing principles of immune signalling, to develop a subsequent generation of vaccine systems for enhanced protection against the most challenging diseases. A New Generation of Protein Interactions: Superglues from Bacteria We have harnessed an amazing feature of the hairs (pili) on the pathogenic bacterium Streptococcus pyogenes. This enabled us to form a spontaneous isopeptide bond between genetically-encoded protein and peptide partners. This interaction is unbreakable, including against high forces in biological systems (blood flow, cell migration, molecular motors). Our favourite pair, SpyTag with SpyCatcher, is one of the strongest protein interactions ever measured. SpyTag is now being applied by hundreds of labs around the world for diverse areas of basic research and biotechnology. We have used computational design and evolution through phage display to create the first genetically-encoded interaction reacting at the diffusion limit and approaching infinite affinity. Through this principle, we have created a series of protein superglues, including SnoopCatcher, DogCatcher and SnoopLigase. We are extending this new class of protein interaction, to create novel possibilities for synthetic biology: • targeting anti-cancer immune responses • SpyRings: cyclised enzymes with extreme resilience, applied for robust diagnostic devices and for enhanced nutrition • protein teams for combinatorial control of cell signalling, towards therapeutics with fewer side-effects. Vaccine development: Unique Protein Architectures for Cell Imaging and Therapy We are re-designing some of the most useful interactions in the biosciences, including antibodies, affibodies and streptavidin: • removing cross-linking (for single molecule imaging of growth factor receptor trafficking) • surpassing one of the strongest non-covalent interactions in nature, for tough diagnostics and mechanical strength (tested at the single molecule level by AFM with Vincent Moy in Miami and by crash-testing DNA pumps) • crystallographic analysis of the limits of non-covalent interaction, creating Love-Hate ligands • hubs for bionanotechnology, enabling assemblies from 4 to 8 to 20 to 180 subunits (maximising T cell detection). Studying the limits of cancer cell capture made clear that even the best antibody interactions are not good enough. We have developed a new class of binding proteins that form covalent bonds to endogenous protein targets. NeissLock was engineered from an adhesion system from Neisseria meningitidis and forms an anhydride in response to calcium, allowing inducibe proximity ligation. Protein ligands that never let go of their targets should reduce the detection limit of soluble biomarkers for early diagnosis, as well as generating long-acting therapeutics and enhancing cell therapy of cancer. Gastrobodies for targeting in the gut The gut is highly effective at degrading proteins. This has prevented the use of antibodies or antibody mimetics for therapeutic targeting in the gut. There are a huge range of bacterial/viral infections, cancers and autoimmune diseases where targeting within the gut could give benefit. We harnessed a protein from soybean with exceptional resistance to the high hydrochloric acid and pepsin concentrations of the stomach. By computational design and evolution, we established a new antibody mimetic called the gastrobody. Gastrobodies could bind and inhibit a C. difficile toxin important for disease progression. We are currently developing the potential of this new targeting platform towards a range of applications in animal and human health. Get in contact for further information about any of these projects, or to discuss the possibility of working on other projects in the area of synthetic biology / vaccines / cancer biology.