Improving Analytical Characterization of Glycoconjugate Vaccines through Combined High-Resolution MS and NMR: Application to Neisseria meningitidis Serogroup B Oligosaccharide-Peptide Glycoconjugates
| Autor: | John F. Cipollo, Yanming An, Marcos D. Battistel, Darón I. Freedberg, Huifeng Yu |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Glycosylation Glycoconjugate Oligosaccharides High resolution Peptide Neisseria meningitidis Serogroup 01 natural sciences Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound Tetanus Toxoid Nuclear Magnetic Resonance Biomolecular Chromatography High Pressure Liquid chemistry.chemical_classification Chromatography Reverse-Phase Vaccines Conjugate 010405 organic chemistry Glycopeptides Nuclear magnetic resonance spectroscopy Oligosaccharide Combinatorial chemistry 0104 chemical sciences Characterization (materials science) 030104 developmental biology chemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Peptides Glycoconjugates Conjugate |
| Zdroj: | Analytical Chemistry. 90:5040-5047 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.7b04748 |
| Popis: | Conjugate vaccines are highly heterogeneous in terms of glycosylation sites and linked oligosaccharide length. Therefore, the characterization of conjugate vaccines' glycosylation state is challenging. However, improved product characterization can lead to enhancements in product control and product quality. Here, we present a synergistic combination of high-resolution mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (NMR) for the analysis of glycoconjugates. We use the power of this strategy to characterize model polysaccharide conjugates and to demonstrate a detailed level of glycoproteomic analysis. These are first steps on model compounds that will help untangle the details of complex product characterization in conjugate vaccines. Ultimately, this strategy can be applied to enhance the characterization of polysaccharide conjugate vaccines. In this study, we lay the groundwork for the analysis of conjugate vaccines. To begin this effort, oligosaccharide-peptide conjugates were synthesized by periodate oxidation of an oligosaccharide of a defined length, α,2-8 sialic acid trimer, followed by a reductive amination, and linking the trimer to an immunogenic peptide from tetanus toxoid. Combined mass spectrometry and nuclear magnetic resonance were used to monitor each reaction and conjugation products. Complete NMR peak assignment and detailed MS information on oxidized oligosialic acid and conjugates are reported. These studies provide a deeper understanding of the conjugation chemistry process and products, which can lead to a better controlled production process. |
| Databáze: | OpenAIRE |
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