Analytical methods play a vital but often hidden role in scientific research. Nuclear magnetic resonance (NMR) spectroscopy is an indispensable technique for probing the structure of molecules at the atomic level and underpins modern drug discovery, industrial process research and studies of biological systems. Most large research institutions have NMR facilities while advances in spectrometer design and automation make the technique fully accessible to non-specialist researchers. Nevertheless, the high purchase and maintenance costs of NMR equipment make time on an NMR spectrometer a precious resource. In the initial period of this project, my team have devised fundamentally new ways of using common equipment that greatly enhance the quantity of information afforded by NMR analysis while shrinking the time and reducing the quantity of sample required. Through innovative experiment design, we can vary the conditions of a sample (pH, solvent composition, molecular concentration) using concentration gradients and interrogate the system using NMR imaging (NMR-I), a 1D variant of magnetic resonance imaging (MRI). By avoiding manual adjustment of the sample conditions, our approach reduces the time required to characterise a chemical system from hours to minutes and enables measurements to be performed that would be unfeasible via conventional approaches. So far, we have demonstrated the rapid assessment of how strongly calcium and magnesium ions bind to food additives (DOI: 10.1021/acs.analchem.2c01166) and how a drug molecule binds to a target protein (DOI: 10.1021/jacs.3c02218). We have also demonstrated the measurement of pH (acidity) in any mixture of solvents, removing the need for cumbersome calibration experiments and liberating experimenters to perform accurate measurements under the exact conditions demanded by their research (DOI: 10.1021/acs.analchem.3c02771). In the later period of this project, we will apply and further develop our methodologies by tackling current problems in biomass processing, microbiology, drug discovery, digestive health and the study of complex molecules such as DNA and enzymes. Four complementary objectives will be pursued, each presenting significant challenges that will require collaboration with partners across other disciplines: Firstly, we will develop a set of tools to identify unknown molecules present in highly complex mixtures. The drive towards sustainable production of products such as pharmaceutical materials has led to the increased use of biomass sources. However, these extracts contain an incredible diversity of molecules, from valuable trace nutrients such as zinc through to large polymers. Identification of these molecules is required for optimisation of extraction processes to maximise yields and minimise waste. The tools we have developed will allow us to identify the functional groups present on the molecules, thus facilitating identification of useful products in biomass extracts and revealing the molecular workings of microorganisms. Secondly, building on our pioneering methods to determine the acidity and protein binding characteristics of model drugs, we will eliminate drawbacks that have emerged during testing of our methodologies with partners in the pharmaceutical industry and further enhance the information afforded in the quest for new drugs. Thirdly, we will develop models of the human gut within standard NMR sample tubes to allow digestive processes to be monitored in real-time, providing a new tool to test fortified foodstuffs. Finally, we will create a new set of tools to evaluate the optimum conditions and rates of activity of enzymes, with potential applications ranging from biotechnology to element cycling within the oceans. We will also explore how the same approach can be used to tailor the rate of formation and properties of DNA secondary structures and self-assembling materials for applications in photovoltaics and wearable electronics.

Modern science is underpinned by efficient and informative analytical methods. Over the past 50 years, nuclear magnetic resonance (NMR) spectroscopy has grown to be one of the dominant analytical techniques in chemical and biological research. A wealth of atomic level information is afforded by NMR on the structure of molecules and their interactions that is inaccessible using other techniques. NMR is vital for the discovery of new drugs, materials and industrial processes and most major research institutions are equipped with NMR facilities. The high purchase and maintenance costs of NMR equipment, along with the widespread utility of the technique, mean that time on an NMR spectrometer is a precious resource. Nevertheless, despite considerable advances in automation, many common procedures involving NMR are extremely demanding in terms of spectrometer time, labour and sample quantity. These demands arise from the frequent requirement to perform multiple NMR measurements on chemical systems as the sample conditions are adjusted (e.g. pH, salt concentration, temperature, solvent composition). For example, the measurement of the pKa value (acidity) of a drug compound requires sets of NMR spectra to be collected as a function of the solution pH. Conventionally, each spectrum must be recorded separately and the pH of the solution adjusted manually between successive NMR experiments. Hours of instrument and analyst time are required to measure this vital property of even a single compound. Similar demands are imposed by the development of temperature or pH-responsive materials for drug delivery systems. The high cost of conventional NMR analysis thus presents a significant barrier to the development of new drugs and materials. In this project, I will create a whole new family of NMR methodologies that will allow the full characterisation of molecular systems in single experiments on single samples with a fraction of the time and cost of conventional approaches. My techniques are based upon NMR imaging (NMR-I), a relative of magnetic resonance imaging (MRI). NMR-I combines the localised analysis afforded by MRI with the wealth of chemical information afforded by NMR. NMR-I can nowadays be performed on almost all NMR equipment without modification and is thus accessible to the majority of researchers. By varying the conditions within a sample and applying NMR-I, it will be possible to perform a full analysis of a system as a function of the sample conditions in just a single experiment. Initial work has shown how, using my methods, 90 individual NMR spectra of a candidate drug molecule can be collected as a function of pH in the time it would take to collect even a single spectrum at a single pH value using conventional approaches. NMR-I will thus accelerate the development and optimisation of new chemical systems while simultaneously freeing up researchers for other duties. There are, however, significant challenges that must be overcome: Firstly, I need to develop ways of creating and analysing controlled gradients of solution properties in standard NMR sample tubes. This is both a theoretical and experimental challenge as little prior work has been done in the field. However, once completed it will be possible to measure the key properties of small molecules, including pharmaceuticals, with unprecedented efficiency. Working with an industrial partner, my methods will be applied to the high-throughput characterisation of compounds in their drug discovery pipeline. Secondly, I will develop techniques that grant researchers access to the novel stimuli-responsive properties of materials such as gels (drug delivery systems, foods, personal care) and polymer electrolytes (DNA, gene vectors, nanotechnology). For example, it will be possible to find the critical conditions at which a drug is released from a binder or a strand of DNA folds. These delicate systems are especially difficult to study using conventional approaches.

Structural biology concerns the study of the three-dimensional structure of biological macromolecules and their interactions. Biomolecular Nuclear Magnetic Resonance (NMR) spectroscopy is one of three core techniques in structural biology, and highly complementary to the other two, i.e. X-ray crystallography and Cryo EM. Extracting information from NMR data has traditionally been complex and non-intuitive. Many smart and innovative tools have been developed, which unfortunately often have been disconnected and not well integrated and sometimes hard to use. For nearly two decades, the Collaborative Computational Project for NMR (CCPN) has been central in providing a connecting interface between the NMR data and many of these tools. CCPN also actively promotes the sharing and exchange of knowledge and best practices. CCPN also actively engages with the UK and international research communities in all matters relating to research funding and policies. The CCPN aims to continue its immense value to the scientific community over the next 5 years by pursuing the following specific objectives: 1. Development of software relevant for NMR CCPN will improve, maintain and expand its programmes, to provide for new functionalities, improved handling, and better speeds. Through fortnightly updates and regular new releases, we will ensure the proper functioning of the software across multiple platforms. We will continuously work on interoperability of our software with other NMR programmes and implement relevant tools for reporting and research data management. 2. NMR in support of Biological Sciences We will facilitate and implement the latest computational tools and developments for NMR data analysis, automation, structure generation and validation. 3. NMR in support of Medicine NMR metabolomics is a thriving field that generates crucial knowledge on metabolic pathways from cells to organisms, including humans. We have designed AnalysisMetabolomics to leverage its power and we will focus on tools for non-expert users, streamlined annotation, assignment, metadata and deposition in public repositories. 4. NMR in support of Industry In collaboration with industrial and academic partners, we will test and enhance applications that are useful to industry. Examples are AnalysisScreen, small-molecule (NMR-assisted) docking procedures to optimise workflows and efficiency and ChemBuild to assist with fragment-based drug discovery. 5. Outreach and training Through its active outreach programme, engaging with all stakeholders including national and major international NMR facilities, CCPN will promote the continuous exchange of knowledge, provide training and support the adoption of best practices in NMR. There is a growing body of "how to" videos available on the website. CCPN will actively continue to promote and develop community data standards (NEF), and will take a leading role in discussions on research funding and policies. CCPN will continue its crucial role as intermediary between the UK and international NMR community, by fostering contacts with (inter-)national NMR facilities, other CCPs, the international wwPDB and NEF efforts. To strengthen the UK NMR community, we will continue the successful series of UK CCPN conferences and teaching programs, our comprehensive help and support for CCPN users, participate in international efforts in knowledge sharing and exchange of best practices, and engage in training and teaching through workshops, papers and (video) tutorials. In short, CCPN will continue to make a crucial difference to the biological NMR community in different fields such as Medicine and Industry as well as its traditional base of the biological sciences by acting as a focal point for technology development, collaboration and sharing best practices. Ultimately, researchers who are empowered with the best tools have more time to make new discoveries.

It is the structural arrangement and motion of molecules and ions that determine, e.g., the bulk properties of a material or the function of biomolecules. The technique of Nuclear Magnetic Resonance (NMR) spectroscopy is very sensitive to the local chemical structure around a particular nucleus, making it a powerful probe of such atomic-level structure and dynamics. To extend the applicability of NMR, two key limiting factors must be addressed: sensitivity, i.e., the relative intensity of spectral peaks as compared to the noise level, and resolution, i.e., the linewidths of individual peaks that determine whether two close-together signals can be separately observed. Both sensitivity and resolution are much improved by performing NMR experiments at higher magnetic field; this proposal is to provide UK researchers with new NMR capability at a world-leading magnetic field strength of 28.2 T, corresponding to a frequency for the 1H nucleus of 1.2 GHz. This builds on the very successful and well-established UK High-Field Solid-State NMR NRF with sustainable ongoing and future operation based on the key factors that have enabled the success of the existing Facility: dedicated Facility Manager support and genuine nationwide buy-in achieved through oversight by a national executive and an independent time allocation procedure. NMR experiments at 28.2 T will make use of as much of the Periodic Table as possible. Nuclei are classified according to their so-called spin quantum number, I. Solution-state NMR on samples is most frequently applied to nuclei with I = 1/2 including such crucial isotopes as 1H, 13C and 15N with correlations between these nuclei traditionally detected on 1H for optimum sensitivity. More recently experiments detected on nuclei other than 1H, especially 13C and 15N, have gained in popularity because of the high resolution achievable for important systems such as intrinsically disordered proteins and large biomolecules including complexes. High field solution NMR is particularly beneficial for biomolecular applications, e.g. characterisation of structures, dynamics and interactions of systems implicated in diseases, but also small molecules, especially for resolving complex mixtures. To maximise the available sensitivity so called cryoprobes, where appropriate parts are kept very cold, are used. In solid-state NMR, the experiment is usually performed by physically rotating the sample around an axis inclined at the so-called magic angle of 54.7 degrees to the magnetic field. For the two most important I = 1/2 nuclei, 1H and 13C, 1.2 GHz will much benefit so-called inverse (i.e., 1H) detection experiments, e.g., for pharmaceuticals and protein complexes, as well as 13C-13C correlation experiments, e.g., for investigating structure and dynamics in plant cell walls. High magnetic field is particularly important for the study of the over two thirds of NMR-active isotopes that possess an electric quadrupole moment, i.e., a non-spherical distribution of electric charge (I of 1 and above). The residual broadening (in the usual NMR scale of ppm) that remains in the magic-angle spinning experiment is inversely proportional to the magnetic field squared; as well as improving resolution, the concentration of the signal intensity into a narrower lineshape means a still greater sensitivity dependence on the magnetic field strength. Application examples include 14N and 35Cl for pharmaceuticals, and 25Mg, 71Ga and 91Zr in materials science. A test of a powerful technique is its applicability to a wide range of problems. The new 1.2 GHz ultra-high magnetic field NMR facility will make possible experiments that provide unique information for applications across science, ranging from materials for catalysis and light harvesting, batteries, drug delivery, to the life sciences, e.g., plant cell walls, protein complexes, membrane proteins and bone structure.