Journal article
Development of an innovative salt-mediated pH gradient cation exchange chromatography method for the characterization of therapeutic antibodies.
- Goyon A School of Pharmaceutical Sciences, University of Geneva, University of Geneva, CMU-Rue Michel Servet 1, 1211 Geneva 4, Switzerland; Institute of Pharmaceutical Sciences of Western Switzerland (ISPSO), University of Geneva, CMU-Rue Michel Servet 1, 1211 Geneva 4, Switzerland.
- McDonald D Protein Analytical Chemistry, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
- Fekete S School of Pharmaceutical Sciences, University of Geneva, University of Geneva, CMU-Rue Michel Servet 1, 1211 Geneva 4, Switzerland; Institute of Pharmaceutical Sciences of Western Switzerland (ISPSO), University of Geneva, CMU-Rue Michel Servet 1, 1211 Geneva 4, Switzerland.
- Guillarme D School of Pharmaceutical Sciences, University of Geneva, University of Geneva, CMU-Rue Michel Servet 1, 1211 Geneva 4, Switzerland; Institute of Pharmaceutical Sciences of Western Switzerland (ISPSO), University of Geneva, CMU-Rue Michel Servet 1, 1211 Geneva 4, Switzerland.
- Stella C Protein Analytical Chemistry, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA. Electronic address: stella.cinzia@gene.com.
- 2020-09-24
Published in:
- Journal of chromatography. B, Analytical technologies in the biomedical and life sciences. - 2020
English
The successful application of monoclonal antibodies (mAb) in oncology and autoimmune diseases paved the way for the development of therapeutic antibodies with a wider range of structural and physico-chemical properties. A pH-gradient combining 2-(N-morpholino)ethanesulfonic (MES) and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffers and mediated with potassium chloride was developed to sufficiently retain acidic mAbs (pI < 7) in cation exchange chromatography (CEX), while keeping suitable separation performance for basic mAbs (pI > 7). Firstly, the MES and HEPES buffers were individually evaluated in their useful pH range by applying a salt gradient. The performance of a salt-mediated pH gradient combining the MES and HEPES buffers was then compared to a commercial pH gradient kit. The developed conditions were found superior to the salt-gradient approaches and provided a useful alternative to commercial pH gradient kits. In this study, the developed conditions were applied to separate a bispecific antibody (BsAb) from its two parental mAbs.
- Language
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- English
- Open access status
- closed
- Identifiers
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- DOI 10.1016/j.jchromb.2020.122379
- PMID 32971367
- Persistent URL
- https://sonar.ch/global/documents/46899
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