Roberto Adamo joined GSK (formerly Novartis Vaccines) in 2007 where he was later appointed as Head of the Carbohydrate Chemistry Laboratory, and Leader of the Conjugation & Synthesis Platform. He is currently Project Leader at GSK, Italy.Luigi Lay is Full Professor at the Department of Chemistry of the University of Milan, Italy. His research focuses on carbohydrate chemistry and the use of synthetic glycans to investigate their biological properties.

• Foreword xvAcknowledgments xvii1 Antibacterial Carbohydrate Vaccines 1Federica Compostella, Laura Morelli, and Luigi Lay1.1 Introduction 11.1.1 A Brief History of Vaccines 21.2 Carbohydrate-Based Vaccines 51.2.1 Mechanism of the Immune Response to Carbohydrate-Based Vaccines 121.3 Components of Glycoconjugate Vaccines 151.3.1 The Carbohydrate Antigen 161.3.2 Linkers for Carbohydrate–Protein Conjugation 191.3.3 The Carrier Protein 221.3.4 The Adjuvant 241.4 Technologies Employed for Production of Glycoconjugate Vaccines 251.4.1 Traditional Glycoconjugates 261.4.2 Glycoconjugates Based on Synthetic Carbohydrate Antigens 281.4.2.1 Site-Selective Protein Conjugation 291.4.3 Enzymatic and ChemoEnzymatic Approach 301.4.4 Bioengineered Glycoconjugates 311.4.5 Nanotechnology-Based Glycoconjugate Vaccines 331.4.5.1 Outer Membrane Vesicles (OMVs) and Generalized Modules for Membrane Antigens (GMMA) 331.4.5.2 Gold Nanoparticles, Liposomes, and Virus-Like Particles 341.4.6 Nonprotein-Based Glycoconjugates 361.4.7 Noncovalent Vaccines 361.5 Conclusion 37Acknowledgments 38References 392 Antifungal Glycoconjugate Vaccines 57Linda del Bino, Maria R. Romano, and Roberto Adamo2.1 Human Fungal Infections 572.2 Immunity Against Fungal Pathogens 592.3 Carbohydrate Antigens in Fungal Cell Wall 602.4 Glycoconjugate Vaccines Against Candida albicans/Candida auris 612.5 Glycoconjugate Vaccines Against Cryptococcus neoformans 642.6 Glycoconjugate Vaccines Against Aspergillus fumigatus 662.7 Universal Fungal Polysaccharide Antigens 682.8 Conclusions and Future Prospects 68References 693 Carbohydrate-Based Antiviral Vaccines 73Adrián Plata and Alberto Fernández-Tejada3.1 Introduction 733.2 Human Immunodeficiency Virus 743.2.1 Vaccine Constructs Derived from gp120 High-Mannose N-Glycan Cluster 753.2.1.1 Surface Oligomannose Cluster-Targeting bnAb: 2G12 Antibody 753.2.1.2 Synthesis and Immunological Evaluation of 2G12 Epitope Mimics 763.2.2 Vaccine Constructs Derived from gp120 First and Second Variable Loops (V1V2) 813.2.2.1 V1V2-Targeting bnAbs 813.2.2.2 Synthetic V1V2 N-Glycopeptide Antigens as bnAb Epitope Mimics 813.2.3 Vaccine Constructs Derived from gp120 Third Variable Loops (V3) 833.2.3.1 V3-Targeting bnAbs 833.2.3.2 Synthetic Glycoconjugates and N-glycopeptides as V3-Directed bnAb Epitope Mimics 833.2.3.3 Synthetic V3 Glycopeptides as bnAb Epitope Mimics 833.3 Influenza A Virus 853.3.1 Vaccine Constructs Based on Hemagglutinin (HA) 863.3.1.1 Hyperglycosylated HA Vaccines 873.3.1.2 α-Gal-Based Vaccine Constructs 873.3.2 Vaccine Constructs Based on Neuraminidase (NA) 883.3.3 Acetalated Dextran as Adjuvant Carrier 893.3.4 Multivalent Constructs as Anti-Influenza Inhibitors 893.4 Hepatitis C Virus 903.5 Ebola Virus 913.5.1 Glycoprotein-Based Vaccines 923.5.2 Monoclonal Antibodies and Carbohydrate Antiviral Agents as Therapeutics 923.6 SARS-CoV-2 Virus 943.6.1 Prospective Vaccine Constructs Based on α-Gal Epitope 943.6.2 RBD-Based Constructs for Vaccine Development 953.6.3 Saponins as Carbohydrate-Based Adjuvant Candidates for COVID- 19 Vaccines 953.7 Conclusions and Outlook 96Acknowledgments 96References 974 Bacterial Glycolipid Lipid As and Their Potential as Adjuvants 111Atsushi Shimoyama and Koichi Fukase4.1 Introduction 1114.2 Bacterial Glycolipid Lipid A: an Innate Immune Stimulant 1134.3 Vaccines Containing Natural LPS as Adjuvants 1174.3.1 Cholera Vaccines 1174.3.2 Salmonella enterica Serovar Typhi Vaccines 1174.3.3 Other Vaccines 1184.4 LPS and Lipid A in the Environment or Fermented Foods as Adjuvants 1184.5 Synthetic and Semisynthetic Lipid As as Adjuvants 1204.6 Developing Novel Lipid A Adjuvants 1214.6.1 Parasitic Bacterial Lipid As 1214.7 Symbiotic Bacterial Lipid As 1234.8 Lipid A-Based Self-Adjuvanting Vaccines 1254.9 Conclusions 127References 1275 Antiadhesive Carbohydrates and Glycomimetics 131Jonathan Cramer, Lijuan Pang, and Beat Ernst5.1 Introduction 1315.1.1 Carbohydrate–Protein Interactions in Viral Adhesion to Host Cells 1315.1.2 Bacterial Adhesins and Antiadhesion Therapy 1325.1.3 Selected Examples 1335.2 DC-SIGN-Mediated Viral Adhesion and Entry into Myeloid Cells 1335.2.1 Introduction 1335.2.2 DC-SIGN Ligands Employing Natural Carbohydrate Epitopes 1365.2.2.1 Dendrimers 1375.2.2.2 Nanoparticles 1375.2.2.3 Polymers 1385.2.2.4 Other Multivalent Scaffolds 1385.2.3 DC-SIGN Ligands Employing Carbohydrate Derivatives or Glycomimetics 1395.2.4 Conclusion and Perspectives 1415.3 The Bacterial Adhesin FimH 1435.3.1 UTIs and FimH 1435.3.2 FimH CRD 1435.3.3 FimH Antagonists 1455.3.4 Conclusion and Perspectives 1475.4 Pseudomonas aeruginosa Virulence Factors (PA-IL and PA-IIL) 1485.4.1 Introduction 1485.4.2 Mono- and Oligovalent Glycomimetic PL-Ligands 1495.4.3 Conclusions and Perspectives 1525.5 General Aspects 152References 1536 Targeting Carbohydrates in Cancer – Analytical and Biotechnological Tools 161Henrique O. Duarte, Joana Gomes, and Celso A. Reis6.1 Aberrant Protein Glycosylation in Cancer 1616.2 Detection and Mapping of Carbohydrate-Based Antigens in Human Neoplastic Tissues 1646.3 Imaging Mass Spectrometry 1646.4 In Situ Proximity Ligation Assay 1666.5 Glycan Microarrays 1696.6 Glycoengineered In Vitro, In Vivo, and Ex Vivo Models 1716.7 Structural Elucidation of Glycoconjugates: Glycomic and Glycoproteomic Strategies 1766.8 Concluding Remarks 182List of Abbreviations 183References 1857 Carbohydrate-Specific Monoclonal Antibody Therapeutics 201Matthew Lohman, Hannah Rowe, and Peter R. Andreana7.1 Introduction 2017.2 Types of Monoclonal Antibodies 2027.2.1 IgG Antibodies 2027.2.2 IgM Antibodies 2037.2.3 ScFv and Fab Fragments 2037.3 Humanization of Monoclonal Antibodies 2047.3.1 CDR Grafting 2047.3.2 Transgenic Animals 2047.4 Breakthrough Research 2057.5 mAbs from Preclinical to Clinical Studies 2067.6 Globo Series 2067.6.1 Blood Group 2067.6.2 Mucin-Attached Glycans 2077.7 New Treatment Options for Neuroblastoma 2077.7.1 History of Unituxin 2087.7.2 What is Unituxin? 2097.7.3 Challenges with Unituxin 2117.7.4 mAbs Binding to Neuroblastoma 2117.7.5 Chimeric and Humanized Anti-GD2 Antibodies 2127.7.6 Naxitamab as a Potential Alternative for High-Risk Patients 2127.7.7 Chimeric Antigen Receptors (CARs) Targeting GD2 2137.8 Summary 214List of Abbreviations 215References 2168 Carbohydrates in Tissue Engineering 223Laura Russo and Francesco Nicotra8.1 Introduction 2238.2 Biomaterials and Medical Devices: Natural and Synthetic Strategies 2248.2.1 Carbohydrates as Building Blocks for Medical Device Formulation 2248.2.1.1 Human Polysaccharides: Glycosaminoglycans (GAGs) and Proteoglycans (PGs) 2258.2.1.2 Polysaccharides from Plants, Algae, Animal, and Microbial Fermentation 2288.2.2 Carbohydrates as Signaling Molecules: Opportunities in Tissue Engineering and Regenerative Medicine 2338.3 Carbohydrates in Animal-Derived Medical Devices: Friends or Foes? 2348.4 Glycoengineering Application to Regenerative Medicine 2358.5 Future Opportunities and Major Challenges 237Conflict of Interest 237References 2379 Carbohydrate-Based Therapeutics for Lysosomal Storage Disorders 245Camilla Matassini, Francesca Clemente, and Francesca Cardona9.1 An Introduction to Lysosomal Storage Disorders (LSDs) 2459.2 Available Treatments for LSDs: The Role of Carbohydrate-Based Therapeutics 2489.2.1 Enzyme Replacement Therapy (ERT) 2509.2.2 Substrate Reduction Therapy (SRT) 2519.2.3 Pharmacological Chaperone Therapy (PCT) 2529.2.4 Combined ERT/PC Therapy 2549.3 Mucopolysaccharidoses 2549.4 Sphingolipidoses 2589.4.1 Fabry Disease 2589.4.2 Gaucher Disease 2629.4.3 Niemann–Pick 2679.4.4 GM1 Gangliosidosis and Morquio B (β-Gal) 2689.4.5 GM2 Gangliosidosis (β-Hexosaminidase) 2729.4.6 Krabbe 2759.5 Glycogen Storage Disorders 2759.5.1 Pompe Disease 2759.6 Glycoproteinoses 2779.6.1 Fucosidosis 2779.6.2 α-Mannosidosis 2799.7 Conclusions 279Acknowledgments 282Abbreviations and Acronyms 283References 28410 Carbohydrates and Carbohydrate-Based Therapeutics in Alzheimer’s Disease 293Ana M. Matos, João Barros, and Amélia P. Rauter10.1 Introduction 29310.2 O-GlcNAc Transferase (OGT) and O-GlcNAc Hydrolase (OGA) in Neurodegeneration 29510.2.1 O-GlcNAc Cycling as a Therapeutic Target Against Alzheimer’s Amyloid Plaques and Neurofibrillary Tangles 29610.2.2 OGA Inhibitors 29910.2.2.1 PUGNAc 30110.2.2.2 GlcNAcstatins 30510.2.2.3 Thiazoline Inhibitors 31110.3 GalNAc in Neurodegeneration 32210.4 Chitosan and Derivatives in AD Brain 32410.5 Cholinesterase Inhibitors 32510.6 Fyn Kinase Inhibitors 33010.7 Amyloid Protein–Protein Interaction Inhibitors 33410.8 Inhibitors of Aβo and/or Oxidative Stress-Induced Neurotoxicity 33810.9 Carbohydrate–Protein Interactions as Potential Therapeutic Targets Against AD 34110.9.1 Lipid-Raft Gangliosides as Membrane Accumulation Sites for Toxic Aβ Aggregates 34110.9.2 The Role of Microglial Cells in Aβ Brain Clearance 34210.10 Conclusion 343List of Abbreviations 344Acknowledgments 347References 34711 Carbohydrate-Based Antithrombotics 353Antonella Bisio, Marco Guerrini, and Annamaria Naggi11.1 Introduction 35311.2 Antithrombotic Drugs 35411.3 Heparin 35411.4 Mechanism of Interaction with Coagulation Factors 35711.4.1 Antithrombin-Mediated Activity 35711.4.2 Heparin Cofactor II Mediated Activity 36011.4.3 Additional Factors 36011.4.4 Adverse Effects of Heparin 36011.4.4.1 Heparin-Induced Thrombocytopenia 36111.4.4.2 Osteoporosis 36111.5 Low Molecular Weight Heparins 36111.5.1 Ultralow Molecular Weight Heparins 36311.6 Drugs Based on Natural GAG Mixtures 36311.6.1 The Role of Dermatan Sulfate 36411.6.2 Sulodexide 36411.6.3 Danaparoid 36511.6.4 Mesoglycan 36511.7 Defibrotide 36611.8 Pentosan Polysulfate 36711.9 Fondaparinux and Related Synthetic Oligosaccharides 36711.10 Chemoenzymatic Synthesis of Oligosaccharides 36911.11 Conclusions and Perspectives 369Acknowledgment 369References 370Index 381

Roberto Adamo joined GSK (formerly Novartis Vaccines) in 2007 where he was later appointed as Head of the Carbohydrate Chemistry Laboratory, and Leader of the Conjugation & Synthesis Platform. He is currently Project Leader at GSK, Italy.Luigi Lay is Full Professor at the Department of Chemistry of the University of Milan, Italy. His research focuses on carbohydrate chemistry and the use of synthetic glycans to investigate their biological properties.

• Foreword xvAcknowledgments xvii1 Antibacterial Carbohydrate Vaccines 1Federica Compostella, Laura Morelli, and Luigi Lay1.1 Introduction 11.1.1 A Brief History of Vaccines 21.2 Carbohydrate-Based Vaccines 51.2.1 Mechanism of the Immune Response to Carbohydrate-Based Vaccines 121.3 Components of Glycoconjugate Vaccines 151.3.1 The Carbohydrate Antigen 161.3.2 Linkers for Carbohydrate–Protein Conjugation 191.3.3 The Carrier Protein 221.3.4 The Adjuvant 241.4 Technologies Employed for Production of Glycoconjugate Vaccines 251.4.1 Traditional Glycoconjugates 261.4.2 Glycoconjugates Based on Synthetic Carbohydrate Antigens 281.4.2.1 Site-Selective Protein Conjugation 291.4.3 Enzymatic and ChemoEnzymatic Approach 301.4.4 Bioengineered Glycoconjugates 311.4.5 Nanotechnology-Based Glycoconjugate Vaccines 331.4.5.1 Outer Membrane Vesicles (OMVs) and Generalized Modules for Membrane Antigens (GMMA) 331.4.5.2 Gold Nanoparticles, Liposomes, and Virus-Like Particles 341.4.6 Nonprotein-Based Glycoconjugates 361.4.7 Noncovalent Vaccines 361.5 Conclusion 37Acknowledgments 38References 392 Antifungal Glycoconjugate Vaccines 57Linda del Bino, Maria R. Romano, and Roberto Adamo2.1 Human Fungal Infections 572.2 Immunity Against Fungal Pathogens 592.3 Carbohydrate Antigens in Fungal Cell Wall 602.4 Glycoconjugate Vaccines Against Candida albicans/Candida auris 612.5 Glycoconjugate Vaccines Against Cryptococcus neoformans 642.6 Glycoconjugate Vaccines Against Aspergillus fumigatus 662.7 Universal Fungal Polysaccharide Antigens 682.8 Conclusions and Future Prospects 68References 693 Carbohydrate-Based Antiviral Vaccines 73Adrián Plata and Alberto Fernández-Tejada3.1 Introduction 733.2 Human Immunodeficiency Virus 743.2.1 Vaccine Constructs Derived from gp120 High-Mannose N-Glycan Cluster 753.2.1.1 Surface Oligomannose Cluster-Targeting bnAb: 2G12 Antibody 753.2.1.2 Synthesis and Immunological Evaluation of 2G12 Epitope Mimics 763.2.2 Vaccine Constructs Derived from gp120 First and Second Variable Loops (V1V2) 813.2.2.1 V1V2-Targeting bnAbs 813.2.2.2 Synthetic V1V2 N-Glycopeptide Antigens as bnAb Epitope Mimics 813.2.3 Vaccine Constructs Derived from gp120 Third Variable Loops (V3) 833.2.3.1 V3-Targeting bnAbs 833.2.3.2 Synthetic Glycoconjugates and N-glycopeptides as V3-Directed bnAb Epitope Mimics 833.2.3.3 Synthetic V3 Glycopeptides as bnAb Epitope Mimics 833.3 Influenza A Virus 853.3.1 Vaccine Constructs Based on Hemagglutinin (HA) 863.3.1.1 Hyperglycosylated HA Vaccines 873.3.1.2 α-Gal-Based Vaccine Constructs 873.3.2 Vaccine Constructs Based on Neuraminidase (NA) 883.3.3 Acetalated Dextran as Adjuvant Carrier 893.3.4 Multivalent Constructs as Anti-Influenza Inhibitors 893.4 Hepatitis C Virus 903.5 Ebola Virus 913.5.1 Glycoprotein-Based Vaccines 923.5.2 Monoclonal Antibodies and Carbohydrate Antiviral Agents as Therapeutics 923.6 SARS-CoV-2 Virus 943.6.1 Prospective Vaccine Constructs Based on α-Gal Epitope 943.6.2 RBD-Based Constructs for Vaccine Development 953.6.3 Saponins as Carbohydrate-Based Adjuvant Candidates for COVID- 19 Vaccines 953.7 Conclusions and Outlook 96Acknowledgments 96References 974 Bacterial Glycolipid Lipid As and Their Potential as Adjuvants 111Atsushi Shimoyama and Koichi Fukase4.1 Introduction 1114.2 Bacterial Glycolipid Lipid A: an Innate Immune Stimulant 1134.3 Vaccines Containing Natural LPS as Adjuvants 1174.3.1 Cholera Vaccines 1174.3.2 Salmonella enterica Serovar Typhi Vaccines 1174.3.3 Other Vaccines 1184.4 LPS and Lipid A in the Environment or Fermented Foods as Adjuvants 1184.5 Synthetic and Semisynthetic Lipid As as Adjuvants 1204.6 Developing Novel Lipid A Adjuvants 1214.6.1 Parasitic Bacterial Lipid As 1214.7 Symbiotic Bacterial Lipid As 1234.8 Lipid A-Based Self-Adjuvanting Vaccines 1254.9 Conclusions 127References 1275 Antiadhesive Carbohydrates and Glycomimetics 131Jonathan Cramer, Lijuan Pang, and Beat Ernst5.1 Introduction 1315.1.1 Carbohydrate–Protein Interactions in Viral Adhesion to Host Cells 1315.1.2 Bacterial Adhesins and Antiadhesion Therapy 1325.1.3 Selected Examples 1335.2 DC-SIGN-Mediated Viral Adhesion and Entry into Myeloid Cells 1335.2.1 Introduction 1335.2.2 DC-SIGN Ligands Employing Natural Carbohydrate Epitopes 1365.2.2.1 Dendrimers 1375.2.2.2 Nanoparticles 1375.2.2.3 Polymers 1385.2.2.4 Other Multivalent Scaffolds 1385.2.3 DC-SIGN Ligands Employing Carbohydrate Derivatives or Glycomimetics 1395.2.4 Conclusion and Perspectives 1415.3 The Bacterial Adhesin FimH 1435.3.1 UTIs and FimH 1435.3.2 FimH CRD 1435.3.3 FimH Antagonists 1455.3.4 Conclusion and Perspectives 1475.4 Pseudomonas aeruginosa Virulence Factors (PA-IL and PA-IIL) 1485.4.1 Introduction 1485.4.2 Mono- and Oligovalent Glycomimetic PL-Ligands 1495.4.3 Conclusions and Perspectives 1525.5 General Aspects 152References 1536 Targeting Carbohydrates in Cancer – Analytical and Biotechnological Tools 161Henrique O. Duarte, Joana Gomes, and Celso A. Reis6.1 Aberrant Protein Glycosylation in Cancer 1616.2 Detection and Mapping of Carbohydrate-Based Antigens in Human Neoplastic Tissues 1646.3 Imaging Mass Spectrometry 1646.4 In Situ Proximity Ligation Assay 1666.5 Glycan Microarrays 1696.6 Glycoengineered In Vitro, In Vivo, and Ex Vivo Models 1716.7 Structural Elucidation of Glycoconjugates: Glycomic and Glycoproteomic Strategies 1766.8 Concluding Remarks 182List of Abbreviations 183References 1857 Carbohydrate-Specific Monoclonal Antibody Therapeutics 201Matthew Lohman, Hannah Rowe, and Peter R. Andreana7.1 Introduction 2017.2 Types of Monoclonal Antibodies 2027.2.1 IgG Antibodies 2027.2.2 IgM Antibodies 2037.2.3 ScFv and Fab Fragments 2037.3 Humanization of Monoclonal Antibodies 2047.3.1 CDR Grafting 2047.3.2 Transgenic Animals 2047.4 Breakthrough Research 2057.5 mAbs from Preclinical to Clinical Studies 2067.6 Globo Series 2067.6.1 Blood Group 2067.6.2 Mucin-Attached Glycans 2077.7 New Treatment Options for Neuroblastoma 2077.7.1 History of Unituxin 2087.7.2 What is Unituxin? 2097.7.3 Challenges with Unituxin 2117.7.4 mAbs Binding to Neuroblastoma 2117.7.5 Chimeric and Humanized Anti-GD2 Antibodies 2127.7.6 Naxitamab as a Potential Alternative for High-Risk Patients 2127.7.7 Chimeric Antigen Receptors (CARs) Targeting GD2 2137.8 Summary 214List of Abbreviations 215References 2168 Carbohydrates in Tissue Engineering 223Laura Russo and Francesco Nicotra8.1 Introduction 2238.2 Biomaterials and Medical Devices: Natural and Synthetic Strategies 2248.2.1 Carbohydrates as Building Blocks for Medical Device Formulation 2248.2.1.1 Human Polysaccharides: Glycosaminoglycans (GAGs) and Proteoglycans (PGs) 2258.2.1.2 Polysaccharides from Plants, Algae, Animal, and Microbial Fermentation 2288.2.2 Carbohydrates as Signaling Molecules: Opportunities in Tissue Engineering and Regenerative Medicine 2338.3 Carbohydrates in Animal-Derived Medical Devices: Friends or Foes? 2348.4 Glycoengineering Application to Regenerative Medicine 2358.5 Future Opportunities and Major Challenges 237Conflict of Interest 237References 2379 Carbohydrate-Based Therapeutics for Lysosomal Storage Disorders 245Camilla Matassini, Francesca Clemente, and Francesca Cardona9.1 An Introduction to Lysosomal Storage Disorders (LSDs) 2459.2 Available Treatments for LSDs: The Role of Carbohydrate-Based Therapeutics 2489.2.1 Enzyme Replacement Therapy (ERT) 2509.2.2 Substrate Reduction Therapy (SRT) 2519.2.3 Pharmacological Chaperone Therapy (PCT) 2529.2.4 Combined ERT/PC Therapy 2549.3 Mucopolysaccharidoses 2549.4 Sphingolipidoses 2589.4.1 Fabry Disease 2589.4.2 Gaucher Disease 2629.4.3 Niemann–Pick 2679.4.4 GM1 Gangliosidosis and Morquio B (β-Gal) 2689.4.5 GM2 Gangliosidosis (β-Hexosaminidase) 2729.4.6 Krabbe 2759.5 Glycogen Storage Disorders 2759.5.1 Pompe Disease 2759.6 Glycoproteinoses 2779.6.1 Fucosidosis 2779.6.2 α-Mannosidosis 2799.7 Conclusions 279Acknowledgments 282Abbreviations and Acronyms 283References 28410 Carbohydrates and Carbohydrate-Based Therapeutics in Alzheimer’s Disease 293Ana M. Matos, João Barros, and Amélia P. Rauter10.1 Introduction 29310.2 O-GlcNAc Transferase (OGT) and O-GlcNAc Hydrolase (OGA) in Neurodegeneration 29510.2.1 O-GlcNAc Cycling as a Therapeutic Target Against Alzheimer’s Amyloid Plaques and Neurofibrillary Tangles 29610.2.2 OGA Inhibitors 29910.2.2.1 PUGNAc 30110.2.2.2 GlcNAcstatins 30510.2.2.3 Thiazoline Inhibitors 31110.3 GalNAc in Neurodegeneration 32210.4 Chitosan and Derivatives in AD Brain 32410.5 Cholinesterase Inhibitors 32510.6 Fyn Kinase Inhibitors 33010.7 Amyloid Protein–Protein Interaction Inhibitors 33410.8 Inhibitors of Aβo and/or Oxidative Stress-Induced Neurotoxicity 33810.9 Carbohydrate–Protein Interactions as Potential Therapeutic Targets Against AD 34110.9.1 Lipid-Raft Gangliosides as Membrane Accumulation Sites for Toxic Aβ Aggregates 34110.9.2 The Role of Microglial Cells in Aβ Brain Clearance 34210.10 Conclusion 343List of Abbreviations 344Acknowledgments 347References 34711 Carbohydrate-Based Antithrombotics 353Antonella Bisio, Marco Guerrini, and Annamaria Naggi11.1 Introduction 35311.2 Antithrombotic Drugs 35411.3 Heparin 35411.4 Mechanism of Interaction with Coagulation Factors 35711.4.1 Antithrombin-Mediated Activity 35711.4.2 Heparin Cofactor II Mediated Activity 36011.4.3 Additional Factors 36011.4.4 Adverse Effects of Heparin 36011.4.4.1 Heparin-Induced Thrombocytopenia 36111.4.4.2 Osteoporosis 36111.5 Low Molecular Weight Heparins 36111.5.1 Ultralow Molecular Weight Heparins 36311.6 Drugs Based on Natural GAG Mixtures 36311.6.1 The Role of Dermatan Sulfate 36411.6.2 Sulodexide 36411.6.3 Danaparoid 36511.6.4 Mesoglycan 36511.7 Defibrotide 36611.8 Pentosan Polysulfate 36711.9 Fondaparinux and Related Synthetic Oligosaccharides 36711.10 Chemoenzymatic Synthesis of Oligosaccharides 36911.11 Conclusions and Perspectives 369Acknowledgment 369References 370Index 381

Roberto Adamo joined GSK (formerly Novartis Vaccines) in 2007 where he was later appointed as Head of the Carbohydrate Chemistry Laboratory, and Leader of the Conjugation & Synthesis Platform. He is currently Project Leader at GSK, Italy.Luigi Lay is Full Professor at the Department of Chemistry of the University of Milan, Italy. His research focuses on carbohydrate chemistry and the use of synthetic glycans to investigate their biological properties.

• Foreword xvAcknowledgments xvii1 Antibacterial Carbohydrate Vaccines 1Federica Compostella, Laura Morelli, and Luigi Lay1.1 Introduction 11.1.1 A Brief History of Vaccines 21.2 Carbohydrate-Based Vaccines 51.2.1 Mechanism of the Immune Response to Carbohydrate-Based Vaccines 121.3 Components of Glycoconjugate Vaccines 151.3.1 The Carbohydrate Antigen 161.3.2 Linkers for Carbohydrate–Protein Conjugation 191.3.3 The Carrier Protein 221.3.4 The Adjuvant 241.4 Technologies Employed for Production of Glycoconjugate Vaccines 251.4.1 Traditional Glycoconjugates 261.4.2 Glycoconjugates Based on Synthetic Carbohydrate Antigens 281.4.2.1 Site-Selective Protein Conjugation 291.4.3 Enzymatic and ChemoEnzymatic Approach 301.4.4 Bioengineered Glycoconjugates 311.4.5 Nanotechnology-Based Glycoconjugate Vaccines 331.4.5.1 Outer Membrane Vesicles (OMVs) and Generalized Modules for Membrane Antigens (GMMA) 331.4.5.2 Gold Nanoparticles, Liposomes, and Virus-Like Particles 341.4.6 Nonprotein-Based Glycoconjugates 361.4.7 Noncovalent Vaccines 361.5 Conclusion 37Acknowledgments 38References 392 Antifungal Glycoconjugate Vaccines 57Linda del Bino, Maria R. Romano, and Roberto Adamo2.1 Human Fungal Infections 572.2 Immunity Against Fungal Pathogens 592.3 Carbohydrate Antigens in Fungal Cell Wall 602.4 Glycoconjugate Vaccines Against Candida albicans/Candida auris 612.5 Glycoconjugate Vaccines Against Cryptococcus neoformans 642.6 Glycoconjugate Vaccines Against Aspergillus fumigatus 662.7 Universal Fungal Polysaccharide Antigens 682.8 Conclusions and Future Prospects 68References 693 Carbohydrate-Based Antiviral Vaccines 73Adrián Plata and Alberto Fernández-Tejada3.1 Introduction 733.2 Human Immunodeficiency Virus 743.2.1 Vaccine Constructs Derived from gp120 High-Mannose N-Glycan Cluster 753.2.1.1 Surface Oligomannose Cluster-Targeting bnAb: 2G12 Antibody 753.2.1.2 Synthesis and Immunological Evaluation of 2G12 Epitope Mimics 763.2.2 Vaccine Constructs Derived from gp120 First and Second Variable Loops (V1V2) 813.2.2.1 V1V2-Targeting bnAbs 813.2.2.2 Synthetic V1V2 N-Glycopeptide Antigens as bnAb Epitope Mimics 813.2.3 Vaccine Constructs Derived from gp120 Third Variable Loops (V3) 833.2.3.1 V3-Targeting bnAbs 833.2.3.2 Synthetic Glycoconjugates and N-glycopeptides as V3-Directed bnAb Epitope Mimics 833.2.3.3 Synthetic V3 Glycopeptides as bnAb Epitope Mimics 833.3 Influenza A Virus 853.3.1 Vaccine Constructs Based on Hemagglutinin (HA) 863.3.1.1 Hyperglycosylated HA Vaccines 873.3.1.2 α-Gal-Based Vaccine Constructs 873.3.2 Vaccine Constructs Based on Neuraminidase (NA) 883.3.3 Acetalated Dextran as Adjuvant Carrier 893.3.4 Multivalent Constructs as Anti-Influenza Inhibitors 893.4 Hepatitis C Virus 903.5 Ebola Virus 913.5.1 Glycoprotein-Based Vaccines 923.5.2 Monoclonal Antibodies and Carbohydrate Antiviral Agents as Therapeutics 923.6 SARS-CoV-2 Virus 943.6.1 Prospective Vaccine Constructs Based on α-Gal Epitope 943.6.2 RBD-Based Constructs for Vaccine Development 953.6.3 Saponins as Carbohydrate-Based Adjuvant Candidates for COVID- 19 Vaccines 953.7 Conclusions and Outlook 96Acknowledgments 96References 974 Bacterial Glycolipid Lipid As and Their Potential as Adjuvants 111Atsushi Shimoyama and Koichi Fukase4.1 Introduction 1114.2 Bacterial Glycolipid Lipid A: an Innate Immune Stimulant 1134.3 Vaccines Containing Natural LPS as Adjuvants 1174.3.1 Cholera Vaccines 1174.3.2 Salmonella enterica Serovar Typhi Vaccines 1174.3.3 Other Vaccines 1184.4 LPS and Lipid A in the Environment or Fermented Foods as Adjuvants 1184.5 Synthetic and Semisynthetic Lipid As as Adjuvants 1204.6 Developing Novel Lipid A Adjuvants 1214.6.1 Parasitic Bacterial Lipid As 1214.7 Symbiotic Bacterial Lipid As 1234.8 Lipid A-Based Self-Adjuvanting Vaccines 1254.9 Conclusions 127References 1275 Antiadhesive Carbohydrates and Glycomimetics 131Jonathan Cramer, Lijuan Pang, and Beat Ernst5.1 Introduction 1315.1.1 Carbohydrate–Protein Interactions in Viral Adhesion to Host Cells 1315.1.2 Bacterial Adhesins and Antiadhesion Therapy 1325.1.3 Selected Examples 1335.2 DC-SIGN-Mediated Viral Adhesion and Entry into Myeloid Cells 1335.2.1 Introduction 1335.2.2 DC-SIGN Ligands Employing Natural Carbohydrate Epitopes 1365.2.2.1 Dendrimers 1375.2.2.2 Nanoparticles 1375.2.2.3 Polymers 1385.2.2.4 Other Multivalent Scaffolds 1385.2.3 DC-SIGN Ligands Employing Carbohydrate Derivatives or Glycomimetics 1395.2.4 Conclusion and Perspectives 1415.3 The Bacterial Adhesin FimH 1435.3.1 UTIs and FimH 1435.3.2 FimH CRD 1435.3.3 FimH Antagonists 1455.3.4 Conclusion and Perspectives 1475.4 Pseudomonas aeruginosa Virulence Factors (PA-IL and PA-IIL) 1485.4.1 Introduction 1485.4.2 Mono- and Oligovalent Glycomimetic PL-Ligands 1495.4.3 Conclusions and Perspectives 1525.5 General Aspects 152References 1536 Targeting Carbohydrates in Cancer – Analytical and Biotechnological Tools 161Henrique O. Duarte, Joana Gomes, and Celso A. Reis6.1 Aberrant Protein Glycosylation in Cancer 1616.2 Detection and Mapping of Carbohydrate-Based Antigens in Human Neoplastic Tissues 1646.3 Imaging Mass Spectrometry 1646.4 In Situ Proximity Ligation Assay 1666.5 Glycan Microarrays 1696.6 Glycoengineered In Vitro, In Vivo, and Ex Vivo Models 1716.7 Structural Elucidation of Glycoconjugates: Glycomic and Glycoproteomic Strategies 1766.8 Concluding Remarks 182List of Abbreviations 183References 1857 Carbohydrate-Specific Monoclonal Antibody Therapeutics 201Matthew Lohman, Hannah Rowe, and Peter R. Andreana7.1 Introduction 2017.2 Types of Monoclonal Antibodies 2027.2.1 IgG Antibodies 2027.2.2 IgM Antibodies 2037.2.3 ScFv and Fab Fragments 2037.3 Humanization of Monoclonal Antibodies 2047.3.1 CDR Grafting 2047.3.2 Transgenic Animals 2047.4 Breakthrough Research 2057.5 mAbs from Preclinical to Clinical Studies 2067.6 Globo Series 2067.6.1 Blood Group 2067.6.2 Mucin-Attached Glycans 2077.7 New Treatment Options for Neuroblastoma 2077.7.1 History of Unituxin 2087.7.2 What is Unituxin? 2097.7.3 Challenges with Unituxin 2117.7.4 mAbs Binding to Neuroblastoma 2117.7.5 Chimeric and Humanized Anti-GD2 Antibodies 2127.7.6 Naxitamab as a Potential Alternative for High-Risk Patients 2127.7.7 Chimeric Antigen Receptors (CARs) Targeting GD2 2137.8 Summary 214List of Abbreviations 215References 2168 Carbohydrates in Tissue Engineering 223Laura Russo and Francesco Nicotra8.1 Introduction 2238.2 Biomaterials and Medical Devices: Natural and Synthetic Strategies 2248.2.1 Carbohydrates as Building Blocks for Medical Device Formulation 2248.2.1.1 Human Polysaccharides: Glycosaminoglycans (GAGs) and Proteoglycans (PGs) 2258.2.1.2 Polysaccharides from Plants, Algae, Animal, and Microbial Fermentation 2288.2.2 Carbohydrates as Signaling Molecules: Opportunities in Tissue Engineering and Regenerative Medicine 2338.3 Carbohydrates in Animal-Derived Medical Devices: Friends or Foes? 2348.4 Glycoengineering Application to Regenerative Medicine 2358.5 Future Opportunities and Major Challenges 237Conflict of Interest 237References 2379 Carbohydrate-Based Therapeutics for Lysosomal Storage Disorders 245Camilla Matassini, Francesca Clemente, and Francesca Cardona9.1 An Introduction to Lysosomal Storage Disorders (LSDs) 2459.2 Available Treatments for LSDs: The Role of Carbohydrate-Based Therapeutics 2489.2.1 Enzyme Replacement Therapy (ERT) 2509.2.2 Substrate Reduction Therapy (SRT) 2519.2.3 Pharmacological Chaperone Therapy (PCT) 2529.2.4 Combined ERT/PC Therapy 2549.3 Mucopolysaccharidoses 2549.4 Sphingolipidoses 2589.4.1 Fabry Disease 2589.4.2 Gaucher Disease 2629.4.3 Niemann–Pick 2679.4.4 GM1 Gangliosidosis and Morquio B (β-Gal) 2689.4.5 GM2 Gangliosidosis (β-Hexosaminidase) 2729.4.6 Krabbe 2759.5 Glycogen Storage Disorders 2759.5.1 Pompe Disease 2759.6 Glycoproteinoses 2779.6.1 Fucosidosis 2779.6.2 α-Mannosidosis 2799.7 Conclusions 279Acknowledgments 282Abbreviations and Acronyms 283References 28410 Carbohydrates and Carbohydrate-Based Therapeutics in Alzheimer’s Disease 293Ana M. Matos, João Barros, and Amélia P. Rauter10.1 Introduction 29310.2 O-GlcNAc Transferase (OGT) and O-GlcNAc Hydrolase (OGA) in Neurodegeneration 29510.2.1 O-GlcNAc Cycling as a Therapeutic Target Against Alzheimer’s Amyloid Plaques and Neurofibrillary Tangles 29610.2.2 OGA Inhibitors 29910.2.2.1 PUGNAc 30110.2.2.2 GlcNAcstatins 30510.2.2.3 Thiazoline Inhibitors 31110.3 GalNAc in Neurodegeneration 32210.4 Chitosan and Derivatives in AD Brain 32410.5 Cholinesterase Inhibitors 32510.6 Fyn Kinase Inhibitors 33010.7 Amyloid Protein–Protein Interaction Inhibitors 33410.8 Inhibitors of Aβo and/or Oxidative Stress-Induced Neurotoxicity 33810.9 Carbohydrate–Protein Interactions as Potential Therapeutic Targets Against AD 34110.9.1 Lipid-Raft Gangliosides as Membrane Accumulation Sites for Toxic Aβ Aggregates 34110.9.2 The Role of Microglial Cells in Aβ Brain Clearance 34210.10 Conclusion 343List of Abbreviations 344Acknowledgments 347References 34711 Carbohydrate-Based Antithrombotics 353Antonella Bisio, Marco Guerrini, and Annamaria Naggi11.1 Introduction 35311.2 Antithrombotic Drugs 35411.3 Heparin 35411.4 Mechanism of Interaction with Coagulation Factors 35711.4.1 Antithrombin-Mediated Activity 35711.4.2 Heparin Cofactor II Mediated Activity 36011.4.3 Additional Factors 36011.4.4 Adverse Effects of Heparin 36011.4.4.1 Heparin-Induced Thrombocytopenia 36111.4.4.2 Osteoporosis 36111.5 Low Molecular Weight Heparins 36111.5.1 Ultralow Molecular Weight Heparins 36311.6 Drugs Based on Natural GAG Mixtures 36311.6.1 The Role of Dermatan Sulfate 36411.6.2 Sulodexide 36411.6.3 Danaparoid 36511.6.4 Mesoglycan 36511.7 Defibrotide 36611.8 Pentosan Polysulfate 36711.9 Fondaparinux and Related Synthetic Oligosaccharides 36711.10 Chemoenzymatic Synthesis of Oligosaccharides 36911.11 Conclusions and Perspectives 369Acknowledgment 369References 370Index 381