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Impact of the column on effluent pH in cation exchange pH gradient chromatography, a practical study.
Farsang, Evelin; Horváth, Krisztián; Beck, Alain; Wang, Qi; Lauber, Matthew; Guillarme, Davy; Fekete, Szabolcs.
Afiliación
  • Farsang E; Department of Analytical Chemistry, University of Pannonia, Egyetem u. 10., H-8200 Veszprém, Hungary.
  • Horváth K; Department of Analytical Chemistry, University of Pannonia, Egyetem u. 10., H-8200 Veszprém, Hungary.
  • Beck A; Center of Immunology Pierre Fabre, 5 Avenue Napoléon III, BP 60497, 74160 Saint-Julien-en-Genevois, France.
  • Wang Q; Waters Corporation, 34 Maple Street, Milford, MA 01757-3696, United States; Current Address: Bristol Myers Squibb, 38 Jackson Rd, Devens, MA 01434, United States.
  • Lauber M; Waters Corporation, 34 Maple Street, Milford, MA 01757-3696, United States.
  • Guillarme D; Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, CMU-Rue Michel Servet 1, 1211 Geneva 4, Switzerland.
  • Fekete S; Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, CMU-Rue Michel Servet 1, 1211 Geneva 4, Switzerland. Electronic address: szabolcs.fekete@unige.ch.
J Chromatogr A ; 1626: 461350, 2020 Aug 30.
Article en En | MEDLINE | ID: mdl-32797830
ABSTRACT
In ionexchange chromatography, the pH gradient mode becomes more and more popular today for the analysis of therapeutic proteins as this mode can provide higher or alternative selectivity to the commonly used salt gradient mode. Ideally, a linear pH response is expected when performing linear gradients. However up to now, only a very few buffer systems have been developed and are commercially available which can perform nearly linear pH responses when flowing through a given column. It is also known that a selected buffer system (mobile phase) can work well on one column but can fail on other column. The goal of this study was to practically evaluate the effects that ionexchange columns (weak and strong exchangers) might have on effluent pH, when performing linear pH gradient separations of therapeutic monoclonal antibodies. To attain this objective, the pH was monitored on-line at the column outlet using a specific setup. To make comprehensive observations of the phenomenon, four different mobile phase conditions and five cation exchange columns (weak and strong exchangers) were employed. The obtained pH responses were systematically compared to responses measured in the absence of the columns. From this work, it has become clear that both the column and mobile phase can have significant effects on pH gradient chromatography and that their combination must be considered when developing a new method. Phase systems (column + mobile phase) providing linear pH responses are indeed the most suitable for separating mAbs with different isoelectric points and, with them, it is possible to elute mAbs across wide retention time ranges and with high selectivity.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Cromatografía por Intercambio Iónico Idioma: En Revista: J Chromatogr A Año: 2020 Tipo del documento: Article País de afiliación: Hungria

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Cromatografía por Intercambio Iónico Idioma: En Revista: J Chromatogr A Año: 2020 Tipo del documento: Article País de afiliación: Hungria
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