Size-exclusion chromatography with multi-angle light scattering for elucidating protein aggregation mechanisms

Abstract

In this chapter, application of size exclusion chromatography with inline multi-angle light scattering (SEC-MALS) to protein systems is reviewed, in particular for its use in elucidating mechanistic details of net-irreversible aggregation processes. After motivating why SEC-MALS or analogous techniques are natural choices to interrogate such aggregating systems, the individual techniques (SEC and MALS) are reviewed briefly, as needed for the context of the remainder of the chapter. Illustrative examples are provided to highlight when and how SEC-MALS can be applied to test mass-action kinetic models for protein aggregation. Limitations of the technique, as well as recommendations for troubleshooting and potential errors in data interpretation are also provided.