Integrated liquid chromatography-multiangle light scattering system to measure absolute masses and stoichiometries of biological assemblies in solution

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This application requests a state-of-the-art size exclusion chromatography system integrated with multi-angle light scattering (SEC-MALS). SEC-MALS is capable of measuring absolute aggregate molecular weights (MWs) and radii of gyration (Rg) under physiological native solution conditions with unparalleled accuracy, precision and throughput not attainable by conventional analytical SEC. Reliable MW and Rg values are needed to determine the oligomerization state and binding stoichiometry of complexes formed by a wide range of proteins and polymers at the center of a multitude of research programs at McMaster. These include not only globular proteins, but also membrane proteins, intrinsically disordered proteins, non-fouling polymers as well as amyloidogenic polypeptides, which are notoriously refractory to other methods for measuring MW and Rg. The requested instrument, a Wyatt MALS-RI with GE Healthcare SEC, is one of the most versatile and reliable SEC-MALS configurations for academic work currently on the market and will have a transformative impact on a collaborative community of > 8 different NSERC-funded research groups spread across three different faculties at McMaster university, i.e. Faculty of Science, Faculty of Health Sciences and Faculty of Engineering. Currently, there is no SEC-MALS at McMaster. Given the outstanding performance in terms of accuracy, sensitivity and throughput, SEC-MALS is now the gold standard requested by the best international journals for the accurate determination of oligomeric states and macromolecular complex stoichiometries. Without SEC-MALS it is becoming nearly impossible to publish high-impact papers. Furthermore, SEC-MALS has been adopted as the industry standard for the characterization of higher-order structures formed by therapeutic biologicals and polymers. Hence, SEC-MALS will dramatically enhance our ability to train the next generation of HQP in preparation for jobs not only in the academia, but also in the pharma and biotech-sectors. The absence of SEC-MALS at McMaster is severely limiting dozens of researchers and is jeopardizing their ability to maintain internationally competitive research programs. If funded, the requested instrument will be the first SEC-MALS in the whole McMaster campus and will be installed in a newly-renovated centralized, shared and widely accessible core facility. We anticipate an extremely high demand by all groups working in the structural biology, enzymology and bioengineering fields. For virtually all projects in these fields, it is imperative to reliably establish the stoichiometry of homo- and hetero-oligomers before further progress can be achieved and currently SEC-MALS is the prime technique for this purpose. The impact of the research enabled by the requested SEC-MALS ranges from the understanding of the mechanisms of allosteric regulation to the design of new biomaterials. Given the absence of SEC-MALS at McMaster, SEC-MALS is a clear and urgent need. **