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Scientific article
English

Theory and practice of size exclusion chromatography for the analysis of protein aggregates

Contributors

Fekete, Szabolcs; Beck, Alain; Veuthey, Jean-Luc; Guillarme, Davy

Published in

Journal of pharmaceutical and biomedical analysis, vol. 101, p. 161-173

Publication date

2014

Abstract

Size exclusion chromatography (SEC) is a historical technique widely employed for the detailed characterization of therapeutic proteins and can be considered as a reference and powerful technique for the qualitative and quantitative evaluation of aggregates. The main advantage of this approach is the mild mobile phase conditions that permit the characterization of proteins with minimal impact on the conformational structure and local environment. Despite the fact that the chromatographic behavior and peak shape are hardly predictable in SEC, some generic rules can be applied for SEC method development, which are described in this review. During recent years, some improvements were introduced to conventional SEC that will also be discussed. Of these new SEC characteristics, we discuss (i) the commercialization of shorter and narrower columns packed with reduced particle sizes allowing an improvement in the resolution and throughput; (ii) the possibility of combining SEC with various detectors, including refractive index (RI), ultraviolet (UV), multi-angle laser light scattering (MALLS) and viscometer (IV), for extensive characterization of protein samples and (iii) the possibility of hyphenating SEC with mass spectrometry (MS) detectors using an adapted mobile phase containing a small proportion of organic modifiers and ion-pairing reagents.

Keywords

  • Size exclusion chromatography
  • Therapeutic proteins
  • Aggregates
  • Biopharmaceuticals

UNIGE affiliation entities

UNIGE research groups

Citation (ISO format)

FEKETE, Szabolcs et al. Theory and practice of size exclusion chromatography for the analysis of protein aggregates. In: Journal of pharmaceutical and biomedical analysis, 2014, vol. 101, p. 161–173. doi: 10.1016/j.jpba.2014.04.011

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Article (Published version)

Identifiers

Additional URL for this publication

http://linkinghub.elsevier.com/retrieve/pii/S0731708514002027

Journal ISSN

0731-7085
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Technical informations

Creation

16/02/2016 16:15:00

First validation

16/02/2016 16:15:00

Update

15/03/2023 00:08:40

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