Advances in size-exclusion separations of proteins and polymers by UHPLC

The separation of molecular compounds based on their capacity to access the intra-particle pore volume of chromatographic media, which is dictated by the relative size in solution of those compounds, has been commonly known as size-exclusion chromatography (SEC) or gel-permeation chromatography (GPC). Conventionally, these two terms have been applied to the analysis of biomolecules and polymers, respectively. Over the more than half-a-century history of size-based separations, there has been a series of advancements, starting from the earliest soft-gel particles and culminating within the past few years in the use of sub-2-µm particles in ultra-high-performance liquid chromatography (UHPLC). The intent of this review is to provide a concise synopsis of the advancements of both chromatography columns and instrumentation for protein and polymer size-based separations. Also, this review presents brief summaries of the application of UHPLC technology for these classes of analytes.