Tuning selectivity in cation-exchange chromatography applied for monoclonal antibody separations, part 2: Evaluation of recent stationary phases

Amarande Murisier, Evelin Farsang, K. Horváth, Matthew Lauber, Alain Beck, Davy Guillarme, Szabolcs Fekete

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

In this second part of the series, recently commercialized cation exchanger stationary phases were systematically investigated for their capabilities to separate therapeutic monoclonal antibodies. It was demonstrated that the different combinations of stationary and mobile phases result in diverse retention, selectivity and efficiency. Hence, the whole phase system (combination of stationary and mobile phase)should be considered when developing a method. In addition, retention behavior is mAb dependent and should be individually optimized. Another interesting observation was that in cation exchange chromatographic separations of large proteins, the particle size of the columns probably impacts retention rather than efficiency, due to the non-porous particle structure - and therefore the higher specific surface area of smaller particles -. Particle size influences the specific surface area and total porosity. Therefore, columns packed with larger particles showed lower retention (when the ion exchanger group was the same e.g. strong exchanger sulfonic group)while no link was observed between efficiency and particle size. The retention, efficiency and selectivity of the studied columns were quite different and strongly dependent on the elution mode (i.e. salt gradient, pH gradient or combined salt/pH gradient mode). The columns can be considered to be complementary, suggesting that it is useful to have more than one type of column on hand while developing new charge variant assays. Moreover, this work shows that it is especially attractive to make use of short, narrow bore ion exchange columns that offer the possibility to perform 4–6 min long separations of both intact and partially digested antibodies.

Original languageEnglish
Pages (from-to)320-328
Number of pages9
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume172
DOIs
Publication statusPublished - Aug 5 2019

Fingerprint

Chromatography
Particle Size
Cations
Ion exchange
Proton-Motive Force
Tuning
Monoclonal Antibodies
Ion exchangers
Particle size
Salts
Specific surface area
Ion Exchange
Porosity
Hand
Observation
Ions
Assays
Antibodies
Proteins
Therapeutics

Keywords

  • Cation exchange chromatography
  • Column efficiency
  • Monoclonal antibody
  • pH gradient
  • Salt gradient
  • Salt-mediated pH gradient

ASJC Scopus subject areas

  • Analytical Chemistry
  • Pharmaceutical Science
  • Drug Discovery
  • Spectroscopy
  • Clinical Biochemistry

Cite this

Tuning selectivity in cation-exchange chromatography applied for monoclonal antibody separations, part 2 : Evaluation of recent stationary phases. / Murisier, Amarande; Farsang, Evelin; Horváth, K.; Lauber, Matthew; Beck, Alain; Guillarme, Davy; Fekete, Szabolcs.

In: Journal of Pharmaceutical and Biomedical Analysis, Vol. 172, 05.08.2019, p. 320-328.

Research output: Contribution to journalArticle

Murisier, Amarande ; Farsang, Evelin ; Horváth, K. ; Lauber, Matthew ; Beck, Alain ; Guillarme, Davy ; Fekete, Szabolcs. / Tuning selectivity in cation-exchange chromatography applied for monoclonal antibody separations, part 2 : Evaluation of recent stationary phases. In: Journal of Pharmaceutical and Biomedical Analysis. 2019 ; Vol. 172. pp. 320-328.
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