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Gregory Staples | Authors

Articles

Troubleshooting LC Separations of Biomolecules, Part 2: Passivation and Mobile-Phase Additives

Bioinert and biocompatible liquid chromatography (LC) systems are becoming more commonplace in laboratories, but the majority of biomolecule separations still use LC systems composed primarily of stainless steel parts. Can passivation or mobile phase additives improve separations on these systems for metal-sensitive biomolecules?

Making the Most of Size‑Exclusion Bioseparations Using Sub‑2‑µm Column Technology

Size-exclusion chromatography (SEC) is a mainstay in the biopharmaceutical industry, serving as a gold standard analytical tool for the characterization of therapeutic proteins in development and manufacturing settings. Contemporary SEC separations can be performed using columns packed with sub-2-μm particles, and these platforms offer the highest efficiencies available for the separation of monoclonal antibody monomer species from low- and high-molecular-weight product-related impurities. Compared to other chromatographic modes used to characterize proteins, SEC is unique in that analytes are not retained by the stationary phase. As a result, special care is required to achieve in practice the chromatographic efficiency that is expected in theory. In this article, we describe the fundamental aspects of achieving high performance using sub-2-μm SEC columns. In addition, we discuss trends in the biopharmaceutical industry, including challenges that can be addressed using modern size-exclusion technologies.