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Characterization of the Conjugation Pattern in Large Polysaccharide–Protein Conjugates by NMR Spectroscopy
Author(s) -
Giuntini Stefano,
Balducci Evita,
Cerofolini Linda,
Ravera Enrico,
Fragai Marco,
Berti Francesco,
Luchinat Claudio
Publication year - 2017
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201709274
Subject(s) - neisseria meningitidis , polysaccharide , glycoconjugate , conjugate , conjugated system , epitope , chemistry , antigen , nuclear magnetic resonance spectroscopy , conjugate vaccine , biochemistry , microbiology and biotechnology , biology , combinatorial chemistry , bacteria , immunogenicity , stereochemistry , immunology , organic chemistry , mathematical analysis , genetics , mathematics , polymer
Abstract Carbohydrate‐based vaccines are among the safest and most effective vaccines and represent potent tools for prevention of life‐threatening bacterial infectious diseases, like meningitis and pneumonia. The chemical conjugation of a weak antigen to protein as a source of T‐cell epitopes generates a glycoconjugate vaccine that results more immunogenic. Several methods have been used so far to characterize the resulting polysaccharide–protein conjugates. However, a reduced number of methodologies has been proposed for measuring the degree of saccharide conjugation at the possible protein sites. Here we show that detailed information on large proteins conjugated with large polysaccharides can be achieved by a combination of solution and solid‐state NMR spectroscopy. As a test case, a large protein assembly, l ‐asparaginase II, has been conjugated with Neisseria meningitidis serogroup C capsular polysaccharide and the pattern and degree of conjugation were determined.