Application Note: Future-proofing the Biopharmaceutical QC Laboratory: Using the ACQUITY UPLC H-Class Bio System to Run SEC-HPLC and SEC-UPLC
9 November 2015
This application note demonstrates the transfer of an SEC-HPLC assay for monoclonal antibody aggregation from legacy HPLC instrumentation to the ACQUITY UPLC H-Class Bio. Aggregation represents a serious concern for companies manufacturing large-molecule therapeutics. Numerous assays aid in establishing the extent of aggregation for a given product, one of which is size exclusion chromatography (SEC). SEC is a straightforward assay requiring minimal sample preparation that exploits the size, or more specifically the hydrodynamic radius, of a given molecule as the mechanism of separation. SEC is unique from many other large molecule chromatographic approaches in the sense that separation occurs under isocratic conditions. Because separation is influenced exclusively by an analyte’s hydrodynamic radius and its ability to penetrate the pores of the stationary phase particle, there is no requirement for gradient conditions as adsorption is ideally non-existent and therefore does not influence migration times.