Roemer, Jennifer and Kiessig, Steffen and Moritz, Bernd and Neusuess, Christian (2021) Improved CE(SDS)-CZE-MS method utilizing an 8-port nanoliter valve. ELECTROPHORESIS, 42 (4). pp. 374-380. ISSN 0173-0835, 1522-2683
Full text not available from this repository. (Request a copy)Abstract
Capillary sieving electrophoresis utilizing SDS (CE(SDS)) is one of the most applied methods for the analysis of antibody (mAb) size heterogeneity in the biopharmaceutical industry. Inadequate peak identification of observed protein fragments is still a major issue. In a recent publication, we introduced an electrophoretic 2D system, enabling online mass spectrometric detection of generic CE(SDS) separated peaks and identification of several mAb fragments. However, an improvement regarding system stability and handling of the approach was desired. Here, we introduce a novel 8-port valve in conjunction with an optimized decomplexation strategy. The valve contains four sample loops with increased distances between the separation dimensions. Thus, successively coinjection of solvent and cationic surfactant without any additional detector in the second dimension is enabled, simplifying the decomplexation strategy. Removal efficiency was optimized by testing different volumes of solvents as presample and cationic surfactant as postsample zone. 2D measurements of the light and heavy chain of the reduced NIST mAb with the 8-port valve and the optimized decomplexation strategy demonstrates the increased robustness of the system. The presented novel set-up is a step toward routine application of CE(SDS)-CZE-MS for impurity characterization of proteins in the biopharmaceutical field.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | CAPILLARY-ELECTROPHORESIS; Capillary electrophoresis; CE(SDS); Mass spectrometry; Monoclonal antibody; Two-dimensional |
| Subjects: | 500 Science > 540 Chemistry & allied sciences |
| Divisions: | Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik |
| Depositing User: | Petra Gürster |
| Date Deposited: | 15 Apr 2021 09:23 |
| Last Modified: | 15 Apr 2021 09:23 |
| URI: | https://pred.uni-regensburg.de/id/eprint/44001 |
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