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

Lead Research Organisation: Imperial College London
Department Name: Chemistry


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, Oxford Nanopore) 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.


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Description The core intention of this award was to provide an SPR facility open to all researchers at Imperial College, our collaborating end-users in industry, and beyond, for the next 10-15 years. The facility has now been operational for several years and is operating at full capacity. Groups from multiple Departments across Imperial College have been trained to use the equipment and have been collecting data for their own research. We anticipate that this will continue for many years, as the instrument helps to provide critical research data, encourages collaborations, and enables new experimental opportunities for sensitive and complex applications.
The Biacore S200 was used in a collaboration with an Imperial startup and has allowed for a robust system to quickly gather kinetic data on target compounds for protein of interest inhibitors, as part of a study in treatment of drug-resistant cancers. A panel of compounds were tested and measured in triplicate with kinetic data collated for comparison to in vitro IC50 measurements. These experiments enabled the selection of highly potent compounds to move forward for further development.
Other research has focussed on the kinetic measurements of compounds that can be inactivated by the application of UV light. The data from these experiments support the theory of a substantial change in binding activity that will help to form a forthcoming publication.
Exploitation Route We envisage the Biacore S200 being operational for a further decade, essentially continuing to deliver high quality kinetic and affinity data that is not possible to gather by other means. These data can inform on the direction of target compounds for research projects in disease areas, or for biotech startups seeking to rapidly identify potential hits for clinical applications. Both Imperial researchers and collaborators will continue to benefit from the SPR facility that has been established for years to come.
Sectors Chemicals,Pharmaceuticals and Medical Biotechnology

Description The Biacore S200 has quickly become an in-demand instrument, available not only to Imperial users at the Molecular Sciences Research Hub (MSRH) at Imperial's White City Campus, but also the wider university. Superusers from the Tate and Barnard groups at the MSRH are running regular training sessions, to enable the waiting list of new users to plan and execute their experiments. Currently, users from seven different research groups across two departments are trained for use on the S200, and we expect this to expand significantly throughout 2020. The current projects underway range from screening of peptides to disrupt protein-protein interactions, assessment of covalent inhibitor binding to protein ligands, and immunoglobulin binding. These experiments will help to rapidly identify the most promising targets for more detailed in vitro characterisations, peptide and probe optimisation, and in cellulo studies for impact in living systems. These findings will substantiate other techniques for publications on cancer related proteins, and bacterial persistence. Users are experimenting with a variety of Series S chips to affix their ligands, standard CM5, NTA, Streptavidin and soon the new Neutravidin chips. The flexibility of approach is enabling users to quickly test their ligands and procure data within a few sessions, whether specificity or equilibrium analysis. We anticipate that the S200 will be critical for producing high quality data for a number of future publications.
First Year Of Impact 2020