en
Scientific article
Review
Open access
English

Unraveling the mysteries of modern size exclusion chromatography – the way to achieve confident characterization of therapeutic proteins

Published inJournal of Chromatography. B, vol. 1092, p. 368-378
Publication date2018
Abstract

Modern size exclusion chromatography (SEC) can be defined by the use of relatively small columns (e.g., 150×4.6mm) packed with sub-3μm particles, allowing a 3- to 5-fold increase in method throughput compared to that of conventional SEC. The quick success of the first sub-2μm SEC column introduced in 2010 led to the development of numerous ultra-high performance (UHP)-SEC columns for the analysis of therapeutic monoclonal antibody (mAb)-based products. Aggregates also known as high-molecular-weight species (HMWS) are indeed one of the most important critical quality attributes (CQAs) of mAbs, as HMWS may decrease the product efficacy or cause immunogenicity effects. Therefore, the confident characterization of mAbs requires strong knowledge of not only modern SEC performance (i.e., selectivity and efficiency) but also the inherent limitations caused by non-specific interactions more likely to occur with complex antibody drug conjugates (ADCs) and some commercial mAb products. This review discusses the importance of liquid chromatographic (LC) instrumentation in order to exploit the full potential of modern SEC columns and current trends to hyphenate SEC to mass spectrometry (MS). Recent applications for antibody-based products (i.e., mAbs, ADCs, Fc-Fusion proteins and bispecific antibodies) are presented. Finally, tips and tricks are provided to further optimize SEC separations and maintaining their performance over time with better understanding of unexpected SEC results.

Keywords
  • Aggregate
  • Antibody drug conjugate
  • Bispecific antibody
  • Biopharmaceutical
  • Fusion proteins
  • Monoclonal antibody
  • Size exclusion chromatography
Citation (ISO format)
GOYON, Alexandre et al. Unraveling the mysteries of modern size exclusion chromatography – the way to achieve confident characterization of therapeutic proteins. In: Journal of Chromatography. B, 2018, vol. 1092, p. 368–378. doi: 10.1016/j.jchromb.2018.06.029
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Identifiers
ISSN of the journal1570-0232
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Technical informations

Creation06/27/2018 2:40:00 PM
First validation06/27/2018 2:40:00 PM
Update time03/15/2023 8:28:30 AM
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