Sugar-Protein Connectivity Impacts on the Immunogenicity of Site-Selective Salmonella O-Antigen Glycoconjugate Vaccines.
A series of glycoconjugates with defined connectivity were synthesized to investigate the impact of coupling Salmonella typhimurium O-antigen to different amino acids of CRM197 protein carrier. In particular, two novel methods for site-selective glycan conjugation were developed to obtain conjugates with single attachment site on the protein, based on chemical modification of a disulfide bond and pH-controlled transglutaminase-catalyzed modification of lysine, respectively. Importantly, conjugation at the C186-201 bond resulted in significantly higher anti O-antigen bactericidal antibody titers than coupling to K37/39, and in comparable titers to conjugates bearing a larger number of saccharides. This study demonstrates that the conjugation site plays a role in determining the immunogenicity in mice and one single attachment point may be sufficient to induce high levels of bactericidal antibodies.
Sclavo Behring Vaccines Institute For Global Health S.r.l., A GSK Company, Via Fiorentina 1, 53100 Siena (Italy).
Giuseppe
G
Stefanetti
Novartis Institutes for BioMedical Research, 100 Technology Square, Cambridge, MA 02139 (USA). qiying.hu@novartis.com.
Qi-Ying
QY
Hu
Novartis Institutes for BioMedical Research, 100 Technology Square, Cambridge, MA 02139 (USA).
Aimee
A
Usera
Novartis Institutes for BioMedical Research, 100 Technology Square, Cambridge, MA 02139 (USA).
Zack
Z
Robinson
Novartis Institutes for BioMedical Research, 100 Technology Square, Cambridge, MA 02139 (USA).
Martin
M
Allan
Novartis Institutes for BioMedical Research, 100 Technology Square, Cambridge, MA 02139 (USA).
Alok
A
Singh
Novartis Institutes for BioMedical Research, 100 Technology Square, Cambridge, MA 02139 (USA).
Hidetomo
H
Imase
Novartis Institutes for BioMedical Research, 100 Technology Square, Cambridge, MA 02139 (USA).
Jennifer
J
Cobb
Novartis Institutes for BioMedical Research, 100 Technology Square, Cambridge, MA 02139 (USA).
Huili
H
Zhai
Novartis Institutes for BioMedical Research, 100 Technology Square, Cambridge, MA 02139 (USA).
Douglas
D
Quinn
Novartis Institutes for BioMedical Research, 100 Technology Square, Cambridge, MA 02139 (USA).
Ming
M
Lei
Sclavo Behring Vaccines Institute For Global Health S.r.l., A GSK Company, Via Fiorentina 1, 53100 Siena (Italy).
Allan
A
Saul
GSK, Via Fiorentina 1, 53100 Siena (Italy).
Roberto
R
Adamo
Jenner Institute, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford, OX3 7DQ (UK).
Calman A
CA
MacLennan
Sclavo Behring Vaccines Institute For Global Health S.r.l., A GSK Company, Via Fiorentina 1, 53100 Siena (Italy). francesca.x.micoli@gsk.com.
Francesca
F
Micoli
D000818:Animals; D005260:Female; D006001:Glycoconjugates / Q000737:chemistry* / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008958:Models, Molecular; D008968:Molecular Conformation; D019081:O Antigens / Q000737:chemistry* / Q000276:immunology*; D022562:Salmonella Vaccines / Q000737:chemistry* / Q000276:immunology*; D012486:Salmonella typhimurium / Q000737:chemistry / Q000276:immunology*
D006001:Glycoconjugates; D019081:O Antigens; D022562:Salmonella Vaccines
carbohydrates; glycoproteins; protein modification; solmonella; vaccines