title: Solution conformation and flexibility of capsular polysaccharides from Neisseria meningitidis and glycoconjugates with the tetanus toxoid protein.
abstract: The structural integrity of meningococcal native, micro-fluidized and activated capsular polysaccharides and their glycoconjugates - in the form most relevant to their potential use as vaccines (dilute solution) - have been investigated with respect to their homogeneity, conformation and flexibility. Sedimentation velocity analysis showed that the polysaccharide size distributions were generally bimodal with some evidence for higher molar mass forms at higher concentration. Weight average molar masses Mwwhere lower for activated polysaccharides. Conjugation with tetanus toxoid protein however greatly increased the molar mass and polydispersity of the final conjugates. Glycoconjugates had an approximately unimodal log-normal but broad and large molar mass profiles, confirmed by sedimentation equilibrium "SEDFIT MSTAR" analysis. Conformation analysis using HYDFIT (which globally combines sedimentation and viscosity data), "Conformation Zoning" and Wales-van Holde approaches showed a high degree of flexibility - at least as great as the unconjugated polysaccharides, and very different from the tetanus toxoid (TT) protein used for the conjugation. As with the recently published finding for Hib-TT complexes, it is the carbohydrate component that dictates the solution behaviour of these glycoconjugates, although the lower intrinsic viscosities suggest some degree of compaction of the carbohydrate chains around the protein.
authors: National Centre for Macromolecular Hydrodynamics, University of Nottingham, Sutton Bonington LE12 5RD, UK.|Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh, 11451, Kingdom of Saudi Arabia.
authors: Ali Saber
authors: AS
authors: Abdelhameed
authors: Department of Chemical Sciences, School of Applied Science, University of Huddersfield, Queensgate, Huddersfield, HD1 3DH, UK.
authors: Gordon A
authors: GA
authors: Morris
authors: Biochemistry Department, Faculty of Science, University of Tabuk, P.O. Box 741-Tabuk 71491 Saudi Arabia.
authors: Fahad
authors: F
authors: Almutairi
authors: National Centre for Macromolecular Hydrodynamics, University of Nottingham, Sutton Bonington LE12 5RD, UK.|Insulin and Diabetes Experimental Research (IDER) Group, University of Nottingham, Faculty of Medicine and Health Science, Clifton Boulevard, Nottingham NG7 2RD, UK.
authors: Gary G
authors: GG
authors: Adams
authors: GSK Vaccines, Rue de l'Institut 89, B1-330 Rixensart, Belgium.
authors: Pierre
authors: P
authors: Duvivier
authors: GSK Vaccines, Rue de l'Institut 89, B1-330 Rixensart, Belgium.
authors: Karel
authors: K
authors: Conrath
authors: National Centre for Macromolecular Hydrodynamics, University of Nottingham, Sutton Bonington LE12 5RD, UK.
authors: Stephen E
authors: SE
authors: Harding
mesh_terms: D000465:Algorithms; D006001:Glycoconjugates / Q000737:chemistry*; D057446:Hydrodynamics; D009345:Neisseria meningitidis / Q000737:chemistry / Q000378:metabolism*; D011135:Polysaccharides, Bacterial / Q000737:chemistry*; D011487:Protein Conformation; D013745:Tetanus Toxoid / Q000737:chemistry*; D014461:Ultracentrifugation
chemical_list: D006001:Glycoconjugates; D011135:Polysaccharides, Bacterial; D013745:Tetanus Toxoid