Enhancing Selectivity of Protein Biopharmaceuticals in Ion Exchange Chromatography through Addition of Organic Modifiers
Charge heterogeneity among therapeutic monoclonal antibodies (mAbs) is considered an important critical quality attribute and requires careful characterization to ensure safe and efficacious drug products. The charge heterogeneity among mAbs is the result of chemical and enzymatic post-translational...
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| Format: | Article |
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MDPI AG
2023-11-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/23/16623 |
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| author | Bastiaan Laurens Duivelshof Thomas Bouvarel Sebastian Pirner Vincent Larraillet Alexander Knaupp Hans Koll Valentina D’Atri Davy Guillarme |
| author_facet | Bastiaan Laurens Duivelshof Thomas Bouvarel Sebastian Pirner Vincent Larraillet Alexander Knaupp Hans Koll Valentina D’Atri Davy Guillarme |
| author_sort | Bastiaan Laurens Duivelshof |
| collection | DOAJ |
| description | Charge heterogeneity among therapeutic monoclonal antibodies (mAbs) is considered an important critical quality attribute and requires careful characterization to ensure safe and efficacious drug products. The charge heterogeneity among mAbs is the result of chemical and enzymatic post-translational modifications and leads to the formation of acidic and basic variants that can be characterized using cation exchange chromatography (CEX). Recently, the use of mass spectrometry-compatible salt-mediated pH gradients has gained increased attention to elute the proteins from the charged stationary phase material. However, with the increasing antibody product complexity, more and more selectivity is required. Therefore, in this study, we set out to improve the selectivity by using a solvent-enriched mobile phase composition for the analysis of a variety of mAbs and bispecific antibody products. It was found that the addition of the solvents to the mobile phase appeared to modify the hydrate shell surrounding the protein and alter the retention behavior of the studied proteins. Therefore, this work demonstrates that the use of solvent-enriched mobile phase composition could be an attractive additional method parameter during method development in CEX. |
| first_indexed | 2024-03-09T01:50:29Z |
| format | Article |
| id | doaj.art-058ad9f8adda431883f800c97660832e |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T01:50:29Z |
| publishDate | 2023-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-058ad9f8adda431883f800c97660832e2023-12-08T15:16:33ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-11-0124231662310.3390/ijms242316623Enhancing Selectivity of Protein Biopharmaceuticals in Ion Exchange Chromatography through Addition of Organic ModifiersBastiaan Laurens Duivelshof0Thomas Bouvarel1Sebastian Pirner2Vincent Larraillet3Alexander Knaupp4Hans Koll5Valentina D’Atri6Davy Guillarme7School of Pharmaceutical Sciences, University of Geneva, CMU—Rue Michel Servet 1, 1211 Geneva, SwitzerlandSchool of Pharmaceutical Sciences, University of Geneva, CMU—Rue Michel Servet 1, 1211 Geneva, SwitzerlandRoche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, GermanyRoche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, GermanyRoche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, GermanyRoche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, GermanySchool of Pharmaceutical Sciences, University of Geneva, CMU—Rue Michel Servet 1, 1211 Geneva, SwitzerlandSchool of Pharmaceutical Sciences, University of Geneva, CMU—Rue Michel Servet 1, 1211 Geneva, SwitzerlandCharge heterogeneity among therapeutic monoclonal antibodies (mAbs) is considered an important critical quality attribute and requires careful characterization to ensure safe and efficacious drug products. The charge heterogeneity among mAbs is the result of chemical and enzymatic post-translational modifications and leads to the formation of acidic and basic variants that can be characterized using cation exchange chromatography (CEX). Recently, the use of mass spectrometry-compatible salt-mediated pH gradients has gained increased attention to elute the proteins from the charged stationary phase material. However, with the increasing antibody product complexity, more and more selectivity is required. Therefore, in this study, we set out to improve the selectivity by using a solvent-enriched mobile phase composition for the analysis of a variety of mAbs and bispecific antibody products. It was found that the addition of the solvents to the mobile phase appeared to modify the hydrate shell surrounding the protein and alter the retention behavior of the studied proteins. Therefore, this work demonstrates that the use of solvent-enriched mobile phase composition could be an attractive additional method parameter during method development in CEX.https://www.mdpi.com/1422-0067/24/23/16623bispecific antibodyFab fragmention exchange chromatographymonoclonal antibodyorganic modifiersalt gradient |
| spellingShingle | Bastiaan Laurens Duivelshof Thomas Bouvarel Sebastian Pirner Vincent Larraillet Alexander Knaupp Hans Koll Valentina D’Atri Davy Guillarme Enhancing Selectivity of Protein Biopharmaceuticals in Ion Exchange Chromatography through Addition of Organic Modifiers International Journal of Molecular Sciences bispecific antibody Fab fragment ion exchange chromatography monoclonal antibody organic modifier salt gradient |
| title | Enhancing Selectivity of Protein Biopharmaceuticals in Ion Exchange Chromatography through Addition of Organic Modifiers |
| title_full | Enhancing Selectivity of Protein Biopharmaceuticals in Ion Exchange Chromatography through Addition of Organic Modifiers |
| title_fullStr | Enhancing Selectivity of Protein Biopharmaceuticals in Ion Exchange Chromatography through Addition of Organic Modifiers |
| title_full_unstemmed | Enhancing Selectivity of Protein Biopharmaceuticals in Ion Exchange Chromatography through Addition of Organic Modifiers |
| title_short | Enhancing Selectivity of Protein Biopharmaceuticals in Ion Exchange Chromatography through Addition of Organic Modifiers |
| title_sort | enhancing selectivity of protein biopharmaceuticals in ion exchange chromatography through addition of organic modifiers |
| topic | bispecific antibody Fab fragment ion exchange chromatography monoclonal antibody organic modifier salt gradient |
| url | https://www.mdpi.com/1422-0067/24/23/16623 |
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