Intact monoclonal antibodies separation and analysis by sheathless capillary electrophoresis-mass spectrometry
| Autor: | Yannis-Nicolas François, Jérémie Giorgetti, Emmanuelle Leize-Wagner, Elise Del Nero, Alain Beck, Antony Lechner |
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| Přispěvatelé: | Laboratoire de spectrométrie de masse des interactions et des systèmes, Chimie de la matière complexe (CMC), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre d'Immunologie Pierre Fabre, Tectonique Moléculaire du Solide (TMS), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA), CIPF, Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de synthèses métallo-induites, Institut de Chimie de Strasbourg-Dynamique et structure moléculaire par spectrométrie de masse (LDSM2) |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Glycosylation
medicine.drug_class 010402 general chemistry Monoclonal antibody Mass spectrometry 01 natural sciences Capillary electrophoresis–mass spectrometry Mass Spectrometry Protein structure Isomerism [CHIM.ANAL]Chemical Sciences/Analytical chemistry Aspartic acid medicine Asparagine Deamidation ComputingMilieux_MISCELLANEOUS Spectroscopy Chromatography Chemistry Biopharmaceutics 010401 analytical chemistry Electrophoresis Capillary General Medicine Trastuzumab Atomic and Molecular Physics and Optics 0104 chemical sciences Protein Processing Post-Translational |
| Zdroj: | European Journal of Mass Spectrometry European Journal of Mass Spectrometry, IM Publications, 2019, 25 (3), pp.324-332. ⟨10.1177/1469066718807798⟩ |
| ISSN: | 1469-0667 |
| DOI: | 10.1177/1469066718807798⟩ |
| Popis: | Capillary electrophoresis mass spectrometry coupling (CE-MS) is a growing technique in biopharmaceutics characterization. Assessment of monoclonal antibodies (mAbs) is well known at middle-up and bottom-up levels to obtain information about the sequence, post-translational modifications (PTMs) and degradation products. Intact protein analysis is an actual challenge to be closer to the real protein structure. At this level, actual techniques are time consuming or cumbersome processes. In this work, a 20 minutes separation method has been developed to optimize characterization of intact mAbs. Thus, separation have been done on a positively-charged coated capillary (PEI) with optimized volatile background electrolyte (BGE) and sample buffer (SB). A sheathless interface allowed to hyphenate CE to a quadrupole-time-of-flight mass spectrometer (Q-TOF) which parameters has been tuned to improve the high masses detection and identification of intact mAbs. Three world-wide health authorities approved mAbs have been used to set up a rapid and ease of use method. Intact trastuzumab, rituximab and palivizumab isoforms have been partially separated with this method in less than 20 minutes under denaturing conditions. For each mAb, 2X-glycosylated and 1X-glycosylated structures have been identified and separated. Concerning basic and acidic variants potential Iso-Asp modification and Asn deamidation have been observed. Accurate mass determination for high-mass molecular species remains a challenge, but the progress in intact mAbs separation appears very promising for biopharmaceutics characterization. |
| Databáze: | OpenAIRE |
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