Comprehensive characterization of bacterial glycoconjugate vaccines by liquid chromatography - mass spectrometry.

Autor: Di Marco F; Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, the Netherlands., Hipgrave Ederveen AL; Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, the Netherlands., van Schaick G; Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, the Netherlands., Moran AB; Bacterial Vaccines Discovery and Early Development, Janssen Vaccines and Prevention B.V., Archimedesweg 4-6, 2333 CN Leiden, the Netherlands., Domínguez-Vega E; Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, the Netherlands., Nicolardi S; Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, the Netherlands., Blöchl C; Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, the Netherlands., Koeleman CA; Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, the Netherlands., Danuser R; Janssen Vaccines AG (Branch of Cilag GmbH International), Rehhagstrasse 79, CH-3018 Bern, Switzerland., Al Kaabi A; Janssen Vaccines AG (Branch of Cilag GmbH International), Rehhagstrasse 79, CH-3018 Bern, Switzerland., Dotz V; Bacterial Vaccines Discovery and Early Development, Janssen Vaccines and Prevention B.V., Archimedesweg 4-6, 2333 CN Leiden, the Netherlands; BioTherapeutics Analytical Development, Janssen Biologics B.V., Einsteinweg 101, 2333 CB Leiden, the Netherlands., Grijpstra J; Bacterial Vaccines Discovery and Early Development, Janssen Vaccines and Prevention B.V., Archimedesweg 4-6, 2333 CN Leiden, the Netherlands., Beurret M; Bacterial Vaccines Discovery and Early Development, Janssen Vaccines and Prevention B.V., Archimedesweg 4-6, 2333 CN Leiden, the Netherlands., Anish C; Bacterial Vaccines Discovery and Early Development, Janssen Vaccines and Prevention B.V., Archimedesweg 4-6, 2333 CN Leiden, the Netherlands., Wuhrer M; Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, the Netherlands. Electronic address: m.wuhrer@lumc.nl.
Jazyk: angličtina
Zdroj: Carbohydrate polymers [Carbohydr Polym] 2024 Oct 01; Vol. 341, pp. 122327. Date of Electronic Publication: 2024 May 30.
DOI: 10.1016/j.carbpol.2024.122327
Abstrakt: Bacterial pathogens can cause a broad range of infections with detrimental effects on health. Vaccine development is essential as multi-drug resistance in bacterial infections is a rising concern. Recombinantly produced proteins carrying O-antigen glycosylation are promising glycoconjugate vaccine candidates to prevent bacterial infections. However, methods for their comprehensive structural characterization are lacking. Here, we present a bottom-up approach for their site-specific characterization, detecting N-glycopeptides by nano reversed-phase liquid chromatography-mass spectrometry (RP-LC-MS). Glycopeptide analyses revealed information on partial site-occupancy and site-specific glycosylation heterogeneity and helped corroborate the polysaccharide structures and their modifications. Bottom-up analysis was complemented by intact glycoprotein analysis using nano RP-LC-MS allowing the fast visualization of the polysaccharide distribution in the intact glycoconjugate. At the glycopeptide level, the model glycoconjugates analyzed showed different repeat unit (RU) distributions that spanned from 1 to 21 RUs attached to each of the different glycosylation sites. Interestingly, the intact glycoprotein analysis displayed a RU distribution ranging from 1 to 28 RUs, showing the predominant species when the different glycopeptide distributions are combined in the intact glycoconjugate. The complete workflow based on LC-MS measurements allows detailed and comprehensive analysis of the glycosylation state of glycoconjugate vaccines.
Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: A.B.M., R.D., V.D., A.A.K., J.G., M.B., and C.A. are employees of Janssen Vaccines. All other authors declare no conflict of interest.
(Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)
Databáze: MEDLINE
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