Ultra-high-performance liquid chromatography for the characterization of therapeutic proteins

The pharmaceutical market has markedly changed over the past few years, and there are today an increasing number of therapeutic drugs produced from biological sources. These biopharmaceuticals include recombinant peptides, proteins, and monoclonal antibodies (mAbs). Their detailed characterization could be difficult and time consuming, so it requires powerful chromatographic and spectroscopic methods. In this context, the use of columns packed with sub-2-µm particles at very high pressure, also known as ultra-high-performance (or pressure) liquid chromatography (UHPLC) has been reported as successful. Various modes of chromatography are compatible with UHPLC columns and conditions, including reversed-phase liquid chromatography (RPLC), size-exclusion chromatography (SEC), ion-exchange chromatography (IEX) and hydrophilic interaction chromatography (HILIC).