title: Click Chemistry Applied to the Synthesis of Salmonella Typhimurium O-Antigen Glycoconjugate Vaccine on Solid Phase with Sugar Recycling.
abstract: A solid-phase conjugation method was developed and applied to the synthesis of an O-antigen based glycoconjugate vaccine against Salmonella Typhimurium, with CRM197 as the carrier protein. Copper-free click chemistry was used as the conjugation chemistry, after derivatizing the sugar and the protein components with alkyne and azido linkers, respectively. This chemistry has the advantage of not deactivating functional groups during the conjugation step, thereby allowing the recycling of unreacted components. The activated carrier protein was adsorbed to an anion exchange matrix and quantitatively conjugated to the O-antigen. The resulting conjugate was eluted from the resin free of unconjugated sugar which was previously removed by simple washing steps. Unreacted O-antigen was recycled by addition to a new batch of resin-CRM197 resulting in further quantitative protein conjugation. This process has advantages in relation to reduction of costs for production of conjugate vaccines, allowing unreacted sugar recovery and simplifying the purification of the glycoconjugate.
authors.affiliation: GSK Vaccines Institute for Global Health (former Novartis Vaccines Institute for Global Health NVGH) , Via Fiorentina 1, 53100 Siena, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Stefanetti
authors.affiliation: GSK Vaccines Institute for Global Health (former Novartis Vaccines Institute for Global Health NVGH) , Via Fiorentina 1, 53100 Siena, Italy.
authors.forename: Allan
authors.initials: A
authors.lastname: Saul
authors.affiliation: Jenner Institute, Nuffield Department of Medicine, University of Oxford , Oxford, OX3 7DQ, United Kingdom.|Wellcome Trust Sanger Institute , Wellcome Trust Genome Campus, Hinxton, Cambridge, CB10 1SA, United Kingdom.
authors.forename: Calman A
authors.initials: CA
authors.lastname: MacLennan
authors.affiliation: GSK Vaccines Institute for Global Health (former Novartis Vaccines Institute for Global Health NVGH) , Via Fiorentina 1, 53100 Siena, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Micoli
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D057930:Click Chemistry / Q000379:methods*; D006001:Glycoconjugates / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D019081:O Antigens / Q000737:chemistry*; D012480:Salmonella Infections / Q000382:microbiology / Q000517:prevention & control; D012486:Salmonella typhimurium / Q000737:chemistry*; D060327:Solid-Phase Synthesis Techniques / Q000379:methods*; D018074:Vaccines, Conjugate / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D006001:Glycoconjugates; D019081:O Antigens; D018074:Vaccines, Conjugate; C028581:CRM197 (non-toxic variant of diphtheria toxin)