Label-free measurement of protein-ligand interactions for biochemistry, structural biology, immunology, engineering, and chemical biology.

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The direct measurement of molecular interactions is essential for understanding proteins and their ligands. Although many tools can provide an estimate in a complex biological milieu using a surrogate measurement (e.g. fluorescent displacement and enzyme-linked assays), others can measure isolated protein-ligand interactions but require labelling (e.g. microscale thermophoresis) or immobilization (e.g. surface plasmon resonance). The former are often label-free but suffer from complications arising from differential protein expression, compound sequestration, and other indirect factors that lead to significant error; the latter are highly accurate, but the modifications of the molecules can affect the validity of the measurement. Isothermal titration calorimetry (ITC) is unique in not requiring any modification of either partner, while directly providing the thermodynamic parameters arising from the binding of two components. Quantifying protein interactions requires significant amounts (mgs) of isolated protein for standard ITC, but the Malvern MicroCal PEAQ-ITC, a nanoITC (nITC) can work with low µg amounts of protein, consistent with the amounts readily available from protein production from our labs. This tool is lacking at UWindsor, at our 2-hours distant closest neighbour, Western, or at our cross-border partners at Wayne State and Oakland University, and will be extensively used by the applicants and frequent users to study a wide range of molecules from biologically active ions, nucleic acids, carbohydrates, peptides and proteins, to supramolecular assemblies such as nanoparticles, viruses, and cavitands and to train our diverse and interdisciplinary next generation of medicinal chemistry and biochemistry researchers. This has also drawn interest from partners at those listed institutions for future collaboration. The nITC is requested by a team of eleven NSERC-funded researchers (two applicants and nine frequent users), including seven early-career investigators across three departments in both science and engineering, spanning the fields of protein science, cellular biology, immunology, drug discovery and novel biomaterials. The researchers conduct ground-breaking innovative science, but the lack of a state-of-the-art instrument capable of studying a wide range of biomolecular interactions hampers the team's research progress. Further delay in the acquisition of this capability has severe implications for HQP training, and will have a long-lasting negative impact on NSERC research programs. This equipment is critical for the training of at least 80 current HQP. The requested instrument is central to the progress of several collaborative, multidisciplinary team grants and industrial contracts that include national and international partnerships in the fields of drug discovery, biotechnology, cellular biology, and the current fight against the COVID-19 pandemic.