Matthew Lauber

This article explores three case studies where autosampler injection programs help to (i) reduce the carry-over observed for large nucleic acids during anion-exchange chromatography (AEX), (ii) reduce peak distortion and breakthrough for monoclonal antibodies (mAbs) during hydrophilic-interaction chromatography (HILIC), and (iii) facilitate dissolution of lipid nanoparticles (LNPs) so that a size-exclusion chromatography (SEC) method can be applied for ribonucleic acid (RNA) payload analysis.

The purpose of this short tutorial article is to review the terms and official nomenclatures for size-exclusion separations and to provide some guidance and recommendations for practicing chromatographers. The interconversion between the different metrics is explained and some examples are presented.

A surge in the development and commercialization of monoclonal antibody (mAb)-based ther-apeutics has driven the requirement for accurate and reproducible analytical methods for pro-tein characterization. Monoclonal antibodies are popular biologic drug candidates, but are sus-ceptible to a myriad of modifications during manufacture and complex degradation pathways during purification and storage, often leading to distinct charge variants that require character-ization and quantification. Ion‑exchange liquid chromatography (IEX) is a well-accepted and widely used technique to separate various mAb charge variant species for the sake of charac-terization and profiling. With the most recent advances in analytical technologies, IEX can be used to help ensure the selection of stable and efficacious mAb drug candidates, from discov-ery through manufacturing.