A surface plasmon resonance facility for label-free analysis of biomolecular interactions

Lead Research Organisation: Imperial College London
Department Name: Dept of Chemistry

Abstract

Life at the molecular level relies on a multitude of intimate, reversible interactions. In the case of macromolecules like proteins, these interactions are responsible for the formation of large multi-subunit complexes, like those that stabilise machineries required to synthesise our genetic material (DNA and RNA polymerases) and all proteins in our body (ribosomes). Other contacts are necessary to stabilise structures that are visible to the naked eye, like the keratin in our hair and nails or the collagen fibres that serve as a flexible scaffold to support different tissues. Some of these interactions are transient, lasting less than a second, whereas others have much longer lifetimes. Determining the strength and the speed of these interactions is essential to identify their biological relevance, e.g. to learn how cells assemble our bodies, generate energy and combat different types of infections. This information is also the basis of important agricultural, biotechnological, medical and industrial applications. For example, modern agrochemical and drug design involves development of a specific ligand (acting as a 'key') that modulates a protein in our cells (acting as a 'lock'). Starting with a poorly binding 'key', cycles of modification with new chemical entities and binding analyses are required to obtain improved versions that recognise a desired 'lock' - leading to compounds that we eventually see as drugs in the market. Surface plasmon resonance (SPR) is currently the gold-standard technique to accurately measure these interactions. Our aim is to replace obsolete equipment at Imperial College London with the latest SPR equipment to allow researchers to accelerate their investigations with this technology.

Technical Summary

Label-free analyses of macromolecular interactions is a critical technology in the biomolecular sciences, playing an essential role in characterisation of complexes in support of structure determination, enzyme mechanism, small molecule ligand or inhibitor identification, membrane protein interactions, antibody and biological therapeutic characterisation, and many other applications. Surface plasmon resonance (SPR) is the gold standard for label-free detection of biomolecular interactions, with unmatched sensitivity and rapid response times capable of measuring binding kinetics and affinities from pico- to millimolar, across size ranges from multi-subunit protein complexes to sub-200 Dalton small molecules.

We propose to establish a new multiuser facility for SPR experiments open to all researchers at Imperial College, our collaborating end-users in industry, and beyond, for the next 10-15 years, through acquisition of a Biacore S200. The Biacore S200 is the world's most sensitive SPR platform, providing the capability to analyse interactions of large, multidomain targets with very low molecular weight compounds, and rare or sensitive targets where only a fraction of the target maintains biological activity during preparation and analysis. The only SPR platform currently at Imperial College is a Biacore 3000 acquired in 1998 and is now obsolete, having been discontinued by the supplier; it suffers from poor sensitivity and low throughput, and lacks small molecule detection capability. The sensitivity and automation capabilities of the new Biacore S200 will thus enable a genuinely transformative range of new applications, from screening of libraries of small molecules (several such libraries are available at Imperial) to studies of proteins with low solubility or limited purification yields, such as membrane proteins.

Planned Impact

We envisage that the largest demand for the facility will come from BBSRC-funded researchers aiming to perform a range of 'single use' experiments of different duration, from individual protein-ligand interactions to screening of large libraries of ligands. These experiments run the full range of BBSRC remit, from structural biology to nutrition, biological mechanisms of aging and antimicrobial resistance. Thus, the multiuser facility will enhance the impact achieved across more than 20 BBSRC-funded projects, programmes and training partnerships, encompassing >£18M BBSRC investment. Through these outputs, the facility will also support BBSRC strategic priorities, delivering impact in collaborative research with users, reducing waste in the food chain, sustainably enhancing agricultural production, bioenergy, synthetic biology, healthy ageing across the lifecourse, combatting antimicrobial resistance, and technology development for the biosciences. The facility will also strongly impact training in the biosciences. Over 200 PhD students, including >100 across the lifecycle of the Imperial BBSRC DTP in Multidisciplinary Training for the Biosciences, and 112 in the EPSRC Centre for Doctoral Training in Chemical Biology.

The scope for new and existing research to leverage this facility for enhanced impact is greatly enhanced by its location in the new £150M Molecular Sciences Research Hub (MSRH) at the Imperial College White City Campus, which co-locates an exciting community of academics, start-ups, spinouts, major international corporations, high-tech and high-growth companies. The MSRH is home to one of the largest chemical biology communities in the world, bringing together the internationally leading Institute of Chemical Biology (ICB) (>135 research groups at Imperial College London), world leading strategic partners in the pharmaceutical (AZ, GSK, Exscientia, Heptares), personal-care (P&G), agri-science (Syngenta), and healthcare/med-tech (NHS, Agilent, OxforNanopore) sectors; key government facilities/institutes (National Physical Laboratory (NPL), Crick Institute, Rosalind Franklin Institute (RFI), Diamond Light Source (DLS); tech accelerators (Rebel Bio); an SME Business Club (>25 members) and >30 affiliated commercial organisations. Space within the MSRH has been specifically set aside for corporations, SMEs, start-ups, spin-outs and entrepreneurs to work alongside Imperial academics, giving our partners unprecedented opportunities for collaboration and access to this multiuser facility. This includes the Imperial College Advanced Hackspace (ICAH), one of the world's largest Hackspaces (>4000 members to date, including members of the general public) providing users with prototyping facilities including robotics, microfluidics, 3-D printing, molecular fabrication, electronics, and synthetic biology.

Besides the MSRH, the new campus houses the Translation & Innovation Hub (I-HUB) and the Michael Uren Biomedical Engineering Research Hub, which from 2019 will house a wide range of research groups from bioengineering, material sciences and medicine. Furthermore, the White City campus is adjacent to Imperial's Hammersmith Hospital which houses a vibrant community of biomedical researchers including the MRC London Institute of Medical Sciences, Imanova Imaging Facility, Imperial Centre for Translational and Experimental Medicine, Imperial Cancer Research UK Centre and Imperial Division of Brain Sciences amongst others.

Publications

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