Project Summary TOC2: High quality and high throughput Small Angle X-ray Scattering A sequence scale structural perspective coupled to an understanding of how these structures change during function is fundamental for mechanistic insights into bio-molecules and can be obtained by combining Small Angle X-ray Scattering (SAXS) with high-resolution snapshots of structure. This strategy has been recently been greatly enabled with the advent of accurate structure prediction from sequence. Furthermore, efficiently identifying the conditions and constructs by SAXS that promote the greatest structural homogeneity in conformation and assembly is hugely advantageous to capture a high-resolution snapshot whether from electron microscopy (EM), macromolecular X-ray crystallography (MX) or nuclear magnetic resonance (NMR). The TOC2 funded SIBYLS group has been instrumental in developing a productive SAXS community and advancing powerful SAXS tools for structural biology. By applying and improving a pipeline that promotes planning, scheduling, executing, analyzing and distributing results for for high-throughput (HT-SAXS) and size exclusion coupled (SEC-SAXS) SAXS, SIBYLS is increasing SAXS utilization by biomedical investigators many-fold. TOC2 at the SIBYLS beamline has explored a unique platform for collecting HT-SAXS data that has had an important impact on protein engineering. The system was designed with a large capacity to increase throughput. Over the course of this renewal we will take advantage of this capacity. TOC2 has also implemented an SEC-SAXS program modelled after the mail-in/hand-in HT-SAXS program. SEC-SAXS is best deployed at synchrotrons as the collection speed matches sample elution speed. SEC-SAXS has had a large impact in the characterization of transient complexes. The increased efficiency and largely remote data collection is particularly important during the pandemic and in the build up to synchrotron shutdowns. Over the next five years through ALS-ENABLE, TOC2 will provide efficient access to SAXS analyses for hundreds of biomedical investigators. The advanced technologies TOC2 employs will generate abundant data that will require analysis by a combination of automated scripts, web applications developed in TOC2 and those developed by peers. We will make use of shared resources and an economy of scale through ALS-ENABLE to support programming of advanced web tools delivering a pipeline that relies heavily on a robust computational infrastructure to organize, process, and store data. By following through on TOC2 aims ALS-ENABLE will increase the value of SAXS experiments and increase accessibility – providing quality in quantity.