Analysis of SEC-SAXS data via EFA deconvolution and Scatter

BioSAXS is a popular technique used in molecular and structural biology to determine the solution structure, particle size shape, surface-to-volume ratio conformational changes of macromolecules macromolecular complexes. A high quality SAXS dataset for modeling must be from monodisperse, homogeneous samples this often only reached by combination inline chromatography immediate measurement. Most commonly, size-exclusion separate exclude contaminants aggregations interest allowing measurements made well-resolved chromatographic peak single protein species. Still, some cases, even purification not guarantee monodisperse samples, either because multiple components are too close each other or shape induced through binding alter perceived elution time. In these it may possible deconvolute data mixture obtain idealized curves individual components. Here, we show how achieved practical analysis SEC-SAXS performed on ideal difficult samples. Specifically, vaccinia E9 DNA polymerase exonuclease minus mutant.