Application Notes: General

The use of sub-2 µm particles for SEC have expanded analytical method for the characterization of biologics. This is especially noteworthy with monoclonal antibodies (mAbs), wherein methods developed on ultra high-performance SEC (UHP-SEC) columns can separate not only aggregates, but also fragment. This application note explores the effect of column internal diameter for robust size exclusion chromatography methods.

Flow rate is a common method parameter for most adsorptive LC methods. However, SEC is unique in that lower flow rates often yield improvements in separation, and resolution can improve considerably simply by running at a lower linear velocity. This application note demonstrates this approach for the aggregate analysis of NIST mAb, a common standard protein.

Assessing phosphate concentration may be necessary to ensure that no differences are observed with modulating concentration. This ensures proper sample recovery and demonstrates robustness of the method. In this application note, we review SEC chromatographic performance for trastuzumab and NIST mAb modulating concentration of potassium phosphate.

Size exclusion chromatography, or SEC, is a technique for separating proteins and other complex biomolecules in their native forms by size using aqueous eluents. For molecules of specific molecular weight, SEC can use used to detect and quantitate aggregates, fragments, monomers and dimer. This guide provides a comprehensive application overview for Biozen dSEC Size Exclusion Columns.