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Research paper thumbnail of Physicochemical studies of a novel adjuvant and conjugate vaccines

CHAPTER 1: Introduction 1.1 Introduction to currently available (paediatric) combination vaccines... more CHAPTER 1: Introduction 1.1 Introduction to currently available (paediatric) combination vaccines CHAPTER 2: Description of physicochemical techniques for the characterisation of adjuvant and vaccine components 2.1 Analysis of Pheroid™ and raw materials by 1H_ and 13C-NMR spectroscopy 74 2.1.1 NMR spectroscopy 74 2.1.2 Size analysis (Coulter Counter) 76 2.2 Analysis of conjugate vaccines 78 2.2.1 Phosphorus determination 78 2.2.2 Orcinol method 2.2.3 High performance anion exchange chromatography (HPAEC) 80 2.2.4 Size exclusion chromatography with ultraviolet detection (SEC-UV) 2.2.5 SCA protein assay 2.3 References CHAPTER 3: Characterisation of Pheroid™ and raw materials by NMR spectroscopy and Coulter Counter analysis 3.1 receptor"" 65 , TolI~like receptors induce signalling pathways, leading to activation of the nuclear factor-KB (NF-KB) in antigen presenting cells, which results in the expression of various cytokine genes, production of co-stimulatory ligands 87-1 (COSO) and 87-2 (COB6), and activation of ada ptive imunity63. 65, It is the initial response of the innate immune system, stimulated by pathogen-associated molecular patterns, that triggers and controls the adaptive immune system response 58 , The binding of pathogenassociated molecular patterns, such as lipopolysaccharide or bacterial CpG-DNA, initiating the activation of adaptive immunity, is indicative of their adjuvant effect acting via specific toll-li ke receptors. Indeed several natural or synthetic adjuvants, including monophosphoryllipid A and lipoarabinomannan , that are bacterial cell•wall components, or the yeast polysaccharide glycan, are structurally closely related to different pathogenassociated molecular pattems S8 • 66. The receptors involved in the interplay of the innate and adaptive immune systems are shown in Fig 1.1. '".,-C\l1ollnn '" Rg 1. 1: Recognition of the pathogen-associated molecular p ettern (PAMP) by pettern-recognition receptors, such as the toll•like receptors, generetes signals that activat6 the adaptive immune sys t&m. ProlfJins dfJrived from the microorganisms ara process6d in the Iysosomes to g6neral6 entig&nic p6ptides, which form a complex with mEtor-histocompatibility-complex (MHC) class /I moIecul6S on fh6 surface of antigen prss6nting cells. Th6se pap/ides are recognized by T-cell receptors. In the case of the signs/ing class of pattern-recognition receptas, the recognition of pathogen-associated molecular patterns by tOlI•llke receptors leads to the activation of signaling pathways that induce the expression of cytCkines, chemCkines, and costimuletory molecules. Therefore, pattern-recognition receptors have a role in the generation of bah the peptide-MHCmolecule complex and the costimulstion mquired for the activation 01 T ce/Is&.

Research paper thumbnail of Physicochemical studies of a novel adjuvant and conjugate vaccines

CHAPTER 1: Introduction 1.1 Introduction to currently available (paediatric) combination vaccines... more CHAPTER 1: Introduction 1.1 Introduction to currently available (paediatric) combination vaccines CHAPTER 2: Description of physicochemical techniques for the characterisation of adjuvant and vaccine components 2.1 Analysis of Pheroid™ and raw materials by 1H_ and 13C-NMR spectroscopy 74 2.1.1 NMR spectroscopy 74 2.1.2 Size analysis (Coulter Counter) 76 2.2 Analysis of conjugate vaccines 78 2.2.1 Phosphorus determination 78 2.2.2 Orcinol method 2.2.3 High performance anion exchange chromatography (HPAEC) 80 2.2.4 Size exclusion chromatography with ultraviolet detection (SEC-UV) 2.2.5 SCA protein assay 2.3 References CHAPTER 3: Characterisation of Pheroid™ and raw materials by NMR spectroscopy and Coulter Counter analysis 3.1 receptor"" 65 , TolI~like receptors induce signalling pathways, leading to activation of the nuclear factor-KB (NF-KB) in antigen presenting cells, which results in the expression of various cytokine genes, production of co-stimulatory ligands 87-1 (COSO) and 87-2 (COB6), and activation of ada ptive imunity63. 65, It is the initial response of the innate immune system, stimulated by pathogen-associated molecular patterns, that triggers and controls the adaptive immune system response 58 , The binding of pathogenassociated molecular patterns, such as lipopolysaccharide or bacterial CpG-DNA, initiating the activation of adaptive immunity, is indicative of their adjuvant effect acting via specific toll-li ke receptors. Indeed several natural or synthetic adjuvants, including monophosphoryllipid A and lipoarabinomannan , that are bacterial cell•wall components, or the yeast polysaccharide glycan, are structurally closely related to different pathogenassociated molecular pattems S8 • 66. The receptors involved in the interplay of the innate and adaptive immune systems are shown in Fig 1.1. '".,-C\l1ollnn '" Rg 1. 1: Recognition of the pathogen-associated molecular p ettern (PAMP) by pettern-recognition receptors, such as the toll•like receptors, generetes signals that activat6 the adaptive immune sys t&m. ProlfJins dfJrived from the microorganisms ara process6d in the Iysosomes to g6neral6 entig&nic p6ptides, which form a complex with mEtor-histocompatibility-complex (MHC) class /I moIecul6S on fh6 surface of antigen prss6nting cells. Th6se pap/ides are recognized by T-cell receptors. In the case of the signs/ing class of pattern-recognition receptas, the recognition of pathogen-associated molecular patterns by tOlI•llke receptors leads to the activation of signaling pathways that induce the expression of cytCkines, chemCkines, and costimuletory molecules. Therefore, pattern-recognition receptors have a role in the generation of bah the peptide-MHCmolecule complex and the costimulstion mquired for the activation 01 T ce/Is&.