A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate

Three important physical properties which may affect the performance of glycoconjugate vaccines against serious disease are molar mass (molecular weight), heterogeneity (polydispersity), and conformational flexibility in solution. The dilute solution behaviour of native and activated capsular polyri...

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Main Authors: Abdelhameed, Ali Saber, Adams, Gary G., Morris, Gordon A., Almutairi, Fahad M., Duvivier, Pierre, Conrath, Karel, Harding, Stephen E.
Format: Article
Published: Nature Publishing Group 2016
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Online Access:https://eprints.nottingham.ac.uk/41697/
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author Abdelhameed, Ali Saber
Adams, Gary G.
Morris, Gordon A.
Almutairi, Fahad M.
Duvivier, Pierre
Conrath, Karel
Harding, Stephen E.
author_facet Abdelhameed, Ali Saber
Adams, Gary G.
Morris, Gordon A.
Almutairi, Fahad M.
Duvivier, Pierre
Conrath, Karel
Harding, Stephen E.
author_sort Abdelhameed, Ali Saber
building Nottingham Research Data Repository
collection Online Access
description Three important physical properties which may affect the performance of glycoconjugate vaccines against serious disease are molar mass (molecular weight), heterogeneity (polydispersity), and conformational flexibility in solution. The dilute solution behaviour of native and activated capsular polyribosylribitol (PRP) polysaccharides extracted from Haemophilus influenzae type b (Hib), and the corresponding glycoconjugate made by conjugating this with the tetanus toxoid (TT) protein have been characterized and compared using a combination of sedimentation equilibrium and sedimentation velocity in the analytical ultracentrifuge with viscometry. The weight average molar mass of the activated material was considerably reduced (Mw ~ 0.24 × 106 g.mol−1) compared to the native (Mw ~ 1.2 × 106 g.mol−1). Conjugation with the TT protein yielded large polydisperse structures (of Mw ~ 7.4 × 106 g.mol−1), but which retained the high degree of flexibility of the native and activated polysaccharide, with frictional ratio, intrinsic viscosity, sedimentation conformation zoning behaviour and persistence length all commensurate with highly flexible coil behaviour and unlike the previously characterised tetanus toxoid protein (slightly extended and hydrodynamically compact structure with an aspect ratio of ~3). This non-protein like behaviour clearly indicates that it is the carbohydrate component which mainly influences the physical behaviour of the glycoconjugate in solution.
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spelling nottingham-416972020-05-04T17:35:20Z https://eprints.nottingham.ac.uk/41697/ A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate Abdelhameed, Ali Saber Adams, Gary G. Morris, Gordon A. Almutairi, Fahad M. Duvivier, Pierre Conrath, Karel Harding, Stephen E. Three important physical properties which may affect the performance of glycoconjugate vaccines against serious disease are molar mass (molecular weight), heterogeneity (polydispersity), and conformational flexibility in solution. The dilute solution behaviour of native and activated capsular polyribosylribitol (PRP) polysaccharides extracted from Haemophilus influenzae type b (Hib), and the corresponding glycoconjugate made by conjugating this with the tetanus toxoid (TT) protein have been characterized and compared using a combination of sedimentation equilibrium and sedimentation velocity in the analytical ultracentrifuge with viscometry. The weight average molar mass of the activated material was considerably reduced (Mw ~ 0.24 × 106 g.mol−1) compared to the native (Mw ~ 1.2 × 106 g.mol−1). Conjugation with the TT protein yielded large polydisperse structures (of Mw ~ 7.4 × 106 g.mol−1), but which retained the high degree of flexibility of the native and activated polysaccharide, with frictional ratio, intrinsic viscosity, sedimentation conformation zoning behaviour and persistence length all commensurate with highly flexible coil behaviour and unlike the previously characterised tetanus toxoid protein (slightly extended and hydrodynamically compact structure with an aspect ratio of ~3). This non-protein like behaviour clearly indicates that it is the carbohydrate component which mainly influences the physical behaviour of the glycoconjugate in solution. Nature Publishing Group 2016-02-26 Article PeerReviewed Abdelhameed, Ali Saber, Adams, Gary G., Morris, Gordon A., Almutairi, Fahad M., Duvivier, Pierre, Conrath, Karel and Harding, Stephen E. (2016) A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate. Scientific Reports, 6 (1). 22208/1-22208/11. ISSN 2045-2322 Biomaterials – vaccines; Glycoconjugates http://www.nature.com/articles/srep22208 doi:10.1038/srep22208 doi:10.1038/srep22208
spellingShingle Biomaterials – vaccines; Glycoconjugates
Abdelhameed, Ali Saber
Adams, Gary G.
Morris, Gordon A.
Almutairi, Fahad M.
Duvivier, Pierre
Conrath, Karel
Harding, Stephen E.
A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate
title A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate
title_full A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate
title_fullStr A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate
title_full_unstemmed A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate
title_short A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate
title_sort glycoconjugate of haemophilus influenzae type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate
topic Biomaterials – vaccines; Glycoconjugates
url https://eprints.nottingham.ac.uk/41697/
https://eprints.nottingham.ac.uk/41697/
https://eprints.nottingham.ac.uk/41697/