title: Conjugation of a peptide to mannan and its confirmation by tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
abstract: The conjugation of polysaccharides to peptides is essential for antigen delivery and vaccine development. Herein, we show that tricine SDS-PAGE in combination with Coomassie Blue staining was adequate to determine the conjugation efficacy of a peptide (epitope 35-55 of myelin oligodendrocyte glycoprotein) to mannan. In addition, tricine SDS-PAGE and periodic acid-Schiff stains were able to monitor the redox state of mannan. Using the described protocol, more than 99.9% of a peptide containing five lysines at its N-terminus was confirmed conjugated to mannan.
authors.affiliation: Department of Chemistry, University of Patras, GR-26504 Rion, Greece.
authors.forename: Anthi
authors.initials: A
authors.lastname: Tapeinou
authors.affiliation: Department of Chemistry, University of Patras, GR-26504 Rion, Greece; Eldrug S.A., Pharmaceutical Company, GR-26504 Platani, Greece.
authors.forename: Maria-Eleni
authors.initials: ME
authors.lastname: Androutsou
authors.affiliation: Department of Chemistry, University of Patras, GR-26504 Rion, Greece.
authors.forename: Konstantina
authors.initials: K
authors.lastname: Kyrtata
authors.affiliation: Department of Chemistry, University of Patras, GR-26504 Rion, Greece. Electronic address: avlamis@upatras.gr.
authors.forename: Alexios
authors.initials: A
authors.lastname: Vlamis-Gardikas
authors.affiliation: Centre for Chronic Disease, College of Health and Biomedicine, Victoria University, Melbourne, VIC, Australia.
authors.forename: Vasso
authors.initials: V
authors.lastname: Apostolopoulos
authors.affiliation: Eldrug S.A., Pharmaceutical Company, GR-26504 Platani, Greece.
authors.forename: John
authors.initials: J
authors.lastname: Matsoukas
authors.affiliation: Department of Chemistry, University of Patras, GR-26504 Rion, Greece. Electronic address: ttselios@upatras.gr.
authors.forename: Theodore
authors.initials: T
authors.lastname: Tselios
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005998:Glycine / Q000031:analogs & derivatives / Q000737:chemistry; D008351:Mannans / Q000737:chemistry*; D063308:Myelin-Oligodendrocyte Glycoprotein / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry
chemical_list: D008351:Mannans; D063308:Myelin-Oligodendrocyte Glycoprotein; D010446:Peptide Fragments; C445081:myelin oligodendrocyte glycoprotein (35-55); D005998:Glycine; C100184:tricine
keywords: Conjugation; MOG(35–55); Mannan; Myelin oligodendrocyte glycoprotein; Peptide; Tricine SDS-PAGE