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Improving Analytical Characterization of Glycoconjugate Vaccines through Combined High-Resolution MS and NMR: Application to Neisseria meningitidis Serogroup B Oligosaccharide-Peptide Glycoconjugates.
- Source :
-
Analytical chemistry [Anal Chem] 2018 Apr 17; Vol. 90 (8), pp. 5040-5047. Date of Electronic Publication: 2018 Apr 03. - Publication Year :
- 2018
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Abstract
- 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.
- Subjects :
- Chromatography, High Pressure Liquid methods
Chromatography, Reverse-Phase
Glycoconjugates chemistry
Glycopeptides analysis
Neisseria meningitidis immunology
Serogroup
Tetanus Toxoid analysis
Tetanus Toxoid chemistry
Vaccines, Conjugate chemistry
Glycoconjugates analysis
Neisseria meningitidis metabolism
Nuclear Magnetic Resonance, Biomolecular
Oligosaccharides chemistry
Peptides chemistry
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods
Vaccines, Conjugate analysis
Subjects
Details
- Language :
- English
- ISSN :
- 1520-6882
- Volume :
- 90
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- Analytical chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 29561588
- Full Text :
- https://doi.org/10.1021/acs.analchem.7b04748