Wyatt Technology Corporation

2 ARTICLES PUBLISHED IN JoVE

Biochemistry

Ion Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization

Hadar Amartely¹, Daniel Some², Ayala Tsadok³, Mario Lebendiker¹

¹Wolfson Centre for Applied Structural Biology, The Alexander Silberman Institute of Life Science, The Hebrew University of Jerusalem, ²Wyatt Technology Corporation, ³Danyel Biotech Ltd

This protocol describes the use of high-specificity ion-exchange chromatography with multi-angle light scattering for an accurate molar mass determination of proteins, protein complexes, and peptides in a heterogeneous sample. This method is valuable for quality assessment, as well as for the characterization of native oligomers, charge variants, and mixed-protein samples.

Biochemistry

Characterization of Proteins by Size-Exclusion Chromatography Coupled to Multi-Angle Light Scattering (SEC-MALS)

Daniel Some¹, Hadar Amartely², Ayala Tsadok³, Mario Lebendiker²

¹Wyatt Technology Corporation, ²Wolfson Centre for Applied Structural Biology, The Alexander Silberman Institute of Life Science, The Hebrew University of Jerusalem, ³Danyel Biotech Ltd.

This protocol describes the combination of size exclusion chromatography with multi-angle light scattering (SEC-MALS) for absolute characterization of proteins and complexes in solution. SEC-MALS determines the molecular weight and size of pure proteins, native oligomers, heterocomplexes and modified proteins such as glycoproteins.