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

该应用程序要求最先进的尺寸排除色谱系统集成多角度光散射（SEC-MALS）。SEC- mals能够测量生理天然溶液条件下的绝对总分子量（MWs）和旋转半径（Rg），具有传统分析SEC无法达到的无与伦比的准确性、精度和吞吐量。在麦克马斯特大学的众多研究项目中，需要可靠的MW和Rg值来确定由各种蛋白质和聚合物形成的复合物的寡聚化状态和结合化学计量学。这些不仅包括球状蛋白，还包括膜蛋白、内在无序蛋白、非污垢聚合物以及淀粉样蛋白多肽，这些都是众所周知的难以用其他方法测量分子量和Rg的。所要求的仪器是带有GE Healthcare SEC的Wyatt MALS-RI，是目前市场上用于学术工作的最通用和最可靠的SEC- mals配置之一，将对分布在麦克马斯特大学三个不同院系（即理学院、健康科学学院和工程学院）的bb80个不同nserc资助的研究小组的协作社区产生变动性影响。目前，麦克马斯特大学没有SEC-MALS。鉴于SEC-MALS在准确性、灵敏度和通量方面的卓越表现，SEC-MALS现已成为国际最佳期刊要求的准确测定低聚物状态和大分子复合物化学计量学的金标准。没有SEC-MALS，发表高影响力的论文几乎是不可能的。此外，SEC-MALS已被采用为表征由治疗性生物制剂和聚合物形成的高阶结构的行业标准。因此，SEC-MALS将大大提高我们培养下一代HQP的能力，不仅为学术界的工作做好准备，也为制药和生物技术部门的工作做好准备。麦克马斯特大学SEC-MALS的缺失严重限制了数十名研究人员的发展，并危及他们维持具有国际竞争力的研究项目的能力。如果获得资助，所要求的仪器将成为整个麦克马斯特校园的第一个SEC-MALS，并将安装在新装修的集中，共享和广泛访问的核心设施中。我们预计在结构生物学，酶学和生物工程领域工作的所有小组都有极高的需求。对于这些领域的几乎所有项目，在取得进一步进展之前，必须可靠地建立同聚物和异聚物的化学计量学，目前SEC-MALS是实现这一目的的主要技术。SEC-MALS的研究影响范围从对变构调节机制的理解到新生物材料的设计。鉴于麦克马斯特大学缺乏SEC-MALS， SEC-MALS是一个明确而迫切的需求。**

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. **