Size exclusion chromatography coupled small angle X-ray scattering with tandem multiangle light scattering at the SIBYLS beamline.

Abstract

Small-angle X-ray Scattering (SAXS) has been a versatile technique for studying biomolecules in solution for several decades now. Developments in SAXS techniques that integrate in situ purification with a high-throughput, multimodal design philosophy have revolutionized the reach and tempo of BioSAXS experiments. The current zenith of the field comes in the form of size exclusion chromatography coupled SAXS with in-line multiangle light scattering (SEC-SAXS-MALS). This technique has been a considerable focus at the Structurally Integrated BiologY for Life Sciences (SIBYLS) beamline at the Advanced Light Source (ALS) in Berkeley, California, over the last 5 years and continues to be a point of active development. In this chapter, we describe the design of the SEC-SAXS-MALS system and general guidelines for collecting, processing, and analyzing SEC-SAXS-MALS data at the SIBYLS beamline.