Benefits of Combined SAXS and UV/Vis Measurements for the Analysis of Biological Samples

Small-angle X-ray scattering (SAXS) and UV/Vis spectroscopy are highly complementary methods for the characterization of e. g., biological molecules in solution. While SAXS probes structural features of the molecules such as size, shape, and folding state, UV/Vis can be used to measure agglomeration and concentration. This report shows the advantages of doing an in-situ combination of both methods probing the exactly same sample volume.

Introduction

Small angle X-ray scattering (SAXS) is a powerful method in structural biology enabling characterization of molecular structure in solution, i.e. in native state. SAXS often fully utilizes its potential in combination with other methods that help interpreting the data or assessing the requirement of particle monodispersity.

Absorption spectroscopy of UV and visible light is such a complementary technique. Typically, it is used for measurements of molecule concentration or moni-toring sample changes (e. g., aggregation) under vari-ous conditions. UV/Vis spectroscopy can be per-formed with very low sample volumes and often in confined space, which makes it ideal for in-situ appli-cations in combination with other methods.

Combining SAXS with in-situ UV/Vis absorption spec-troscopy enables the characterization of samples us-ing both methods on the same sample volume at the same time. As an example, this report highlights con-centration measurement and temperature induced aggregation monitoring of a protein sample.

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