Proteins, monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs) are powerful therapeutic agents. Because of their high molecular complexity, however, biopharmaceuticals may be quite sensitive to changes in the manufacturing processes that can lead to considerable micro-heterogeneity. As a result, a panel of separation techniques based on both liquid chromatography and electrophoresis has been used for their characterization and comparability studies. In terms of detection, mass spectrometry (MS) plays a pivotal role in the structural elucidation of biopharmaceuticals, because it offers an additional degree of separation by mass/charge ratio, greatly facilitating the characterization of variants. This two-part web seminar will discuss possibilities and limitations of chromatographic techniques and mass spectrometry detection for the physico-chemical characterization of biopharmaceutical compounds. Part I will focus on chromatographic techniques used for the physico-chemical characterization of biopharmaceutical compounds. Historically, ion exchange chromatography (IEX) and size exclusion chromatography (SEC) were the reference techniques. In this presentation, some new trends in IEX and SEC will be explained. However, because reversed-phase liquid chromatography is naturally more compatible with MS, the talk will focus on the potential of state-of-the-art UHPLC instruments and reversed-phase LC columns (sub-2?m wide-pore porous particles or core-shell particles) for biopharmaceutical characterization.
