A Review Size-Exclusion Chromatography for the Analysis of Protein Biotherapeutics and Their Aggregates

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Paula Hong, Stephan Koza, and Edouard S. P. Bouvier
Journal of Liquid Chromatography & Related Technologies,
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Scientific Publications
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In recent years, the use and number of biotherapeutics has increased significantly. For these largely protein-based therapies, the quantitation of aggregates is of particular concern given their potential effect on efficacy and immunogenicity. This need has renewed interest in size-exclusion chromatography (SEC). In the following review we will outline the history and background of SEC for the analysis of proteins. We will also discuss the instrumentation for these analyses, including the use of different types of detectors. Method development for protein analysis by SEC will also be outlined, including the effect of mobile phase and column parameters (column length, pore size). We will also review some of the applications of this mode of separation that are of particular importance to protein biopharmaceutical development and highlight some considerations in their implementation.

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