Aggregation of proteins is a commonplace phenomenon and a serious danger to safety and effectiveness of medical products. It is the obligation of each manufacturer to control the level of non-native structures of proteins. One of the methods that can be applied in such analyses is exclusion chromatography combined with the MALS (SEC-MALS) detector. This technique allows separating individual fractions in accordance with their hydrodynamic radius and then establishing the radius of gyration as well as the weight of particles representing individual fractions.
We have the HPLC Dionex ICS 5000+ system equipped with the UV detector, autosampler with the option of a thermostat which is combined with the mini Dawn Treos MALS by Wyatt Technology. These systems allow us to analyze the particles with the weight of 200 Da up to 10 MDa and the radius of gyration ranging from 10 nm up to 50 nm.
Given the application of the appropriate columns and mobile phases we can differentiate the protein aggregates from the properly formed oligomers.