Absolute Molar Mass in UHPLC via the μDAWN and UT-rEX

Size exclusion chromatography (SEC) is widely used for characterizing the size and composition of aggregates or fragments in a biomolecular sample. Conventional SEC relies on reference standards to calibrate column elution time as a function of molar mass. However, these standards often do not accurately represent the conformation of the sample, or the sample may exhibit column interactions that change its elution properties relative to the standard. This application note demonstrates the coupling of a light scattering detector, such as the miniDAWN™ TREOS®, to SEC, enabling absolute quantification of a molecule’s molar mass, irrespective of elution time.