| Added by | pmartino |
|---|---|
| Group name | EquipePM |
| Item Type | Journal Article |
| Title | An in-line enzymatic microreactor for the middle-up analysis of monoclonal antibodies by capillary electrophoresis |
| Creator | Dadouch et al. |
| Author | Meriem Dadouch |
| Author | Yoann Ladner |
| Author | Claudia Bich |
| Author | Marion Larroque |
| Author | Christian Larroque |
| Author | Jacques Morel |
| Author | Pierre-Antoine Bonnet |
| Author | Catherine Perrin |
| Abstract | Monoclonal antibodies (mAbs) are undergoing rapid growth in the pharmaceutical industry due to their clinical efficiency. Concomitantly, robust, cost-effective, and high throughput analytical methods are needed for their quality control. Among all analytical techniques, capillary electrophoresis (CE) presents alternative and attractive features because the capillary can be used both as a microreactor and as a support for separation. Transverse diffusion of laminar flow profiles was applied for the middle-up analysis of mAbs for the first time. Infliximab was selected as the model mAb. All middle-up analysis steps (enzymatic digestion, electrophoretic separation and UV detection) were integrated into the same capillary. The conditions for the separation of infliximab subunits (pH, ionic strength, and type of background electrolyte) and in-line digestion parameters (reactant injection conditions, time, temperature and enzyme/mAb ratio) were optimized. The in-line methodology was compared to the off-line methodology and evaluated in terms of proteolysis efficiency, repeatability, and applicability to different mAbs. Finally, the methodology was transferred to capillary electrophoresis coupled to mass spectrometry (sheathless interface) to identify infliximab subunits. The in-line methodology was successfully implemented with a simplified injection scheme, temperature control, fast enzymatic reaction and high resolution of separation of infliximab subunits under pseudo-native MS compatible conditions. In comparison with the off-line methodology, reactant consumption was reduced by a factor of 1000, and the numbers of theoretical plates were increased by a factor of 2. |
| Publication | The Analyst |
| Date | Jan 08, 2020 |
| Journal Abbr | Analyst |
| Language | eng |
| DOI | 10.1039/c9an01906e |
| ISSN | 1364-5528 |
| Library Catalog | PubMed |
| Extra | 00000 PMID: 31913378 |
| Tags | original |
| Date Added | 2020/02/07 - 08:23:57 |
| Date Modified | 2021/03/05 - 10:42:44 |
| Notes and Attachments | PubMed entry (Attachment) PubMed entry (Attachment) |
Institut de Recherche en Cancérologie de
