Capillary Electrophoresis of Carbohydrates (eBook)

Preface 6
Contents 10
Contributors 12
Chapter 1: Analysis of Simple Carbohydrates by Capillary Electrophoresis and Capillary Electrophoresis–Mass Spectrometry 16
1.1 Introduction 17
1.2 Determination of Simple Carbohydrates by CE 17
1.2.1 Detection Systems in CE 24
1.2.1.1 Spectrophotometric Detection for the Determination of Underivatized Carbohydrates by CE 24
1.2.1.2 Electrochemical Detection for the Determination of Carbohydrates by CE 27
1.2.1.3 Other Detection Methods for the Determination of Carbohydrates by CE 29
1.2.2 Electrolyte Systems for the CE-Separation of Simple Carbohydrates 30
1.2.3 Analysis of Simple Carbohydrates in Real Samples by CE 32
References 32
Chapter 2: Fluorophores and Chromophores for the Separation of Carbohydrates by Capillary Electrophoresis 37
2.1 Introduction 38
2.2 Reductive Amination 40
2.3 Amination Via Formation of Glycosylamine 53
2.4 Derivatization with N-Methyl-glycamine Derivatives 53
2.5 Formation of Hydrazones 55
2.6 Derivatization with Pyrazolone Compounds 56
2.7 Derivatization at Carboxylic Acid Functionalities 58
2.8 On-Column Derivatization 59
2.9 Conclusion and Future Directions 61
References 61
Chapter 3: Capillary Electrophoresis of Bacterial (Lipo)Polysaccharides 66
3.1 Introduction 68
3.2 Capillary Electrophoresis of Lipopolysaccharides 69
3.2.1 On-Gel Hydrolysis and CE-LIF Monosaccharide Composition of Excised Lipopolysaccharides 71
3.2.2 CE Chips for Screening of Endotoxin Chemotypes from Whole-Cell Lysates 72
3.2.3 Capillary Zone Electrophoresis (CZE) Separation of O-Deacylated LPS and Polysaccharide Derivatives 74
3.2.4 CE for Quantitative Analysis of Extracted Lipopolysaccharides 75
3.2.5 CZE for the Determination of Meningococcal Polysaccharides 75
3.2.6 CE to Monitor Lipopolysaccharides (Endotoxins) by Protein Complexation 78
3.2.7 CE of Alginic Acid 80
3.2.8 Characterization of Lipopolysaccharides by CE-MS 82
3.2.8.1 Characterization of Intact LPS from the H. influenzae by Electrophoresis-Assisted Open-Tubular Liquid Chromatography-MS 82
3.2.8.2 Carbohydrate Analysis of Typical and Atypical Isolates of A. salmonicida for the LPS-Based Classification 83
3.2.8.3 CE and High-Field Asymmetric Waveform Ion Mobility Spectrometry MS for the Analysis of LPS 84
3.2.8.4 Glycotyping by CE-MS of Pseudomonas aeruginosa Isolates from Patients with Cystic Fibrosis 85
3.2.8.5 Characterization of Short-Chain LPS Glycoforms by CE-MS 86
3.2.8.6 Isomer Separation and Characterization by CE-MS 86
3.2.8.7 In-Source Fragmentation Strategy for Polysaccharide Analysis 87
3.3 CE Analysis of Bacterial Glycosaminoglycan-Like Polysaccharides 87
3.3.1 General Considerations 87
3.3.2 CE Analysis of E. coli K4 Polysaccharide 88
3.3.3 CE Analysis of E. coli K5 Polysaccharide (Heparosan) 90
3.4 Conclusion 91
References 92
Chapter 4: Capillary Electrophoresis and Its Microchip Format for the Analysis of Glycosaminoglycans 96
4.1 Introduction 97
4.2 CE Analysis of GAGs 99
4.2.1 Analysis of Hyaluronic Acid and Its Oligosaccharides 99
4.2.2 Analysis of Sulfated GAGs 101
4.2.2.1 Intact Sulfated GAGs 101
4.2.2.2 Unsaturated Disaccharides 103
4.3 Microchip Electrophoresis of GAGs 104
4.3.1 Analysis of Di- and Oligosaccharides 106
4.3.2 Analysis of GAG Polysaccharides 109
4.4 Conclusion 112
References 113
Chapter 5: Extracellular Polysaccharides in Microbial Biofilm and Their Influence on the Electrophoretic Properties of Microbial Cells 117
5.1 Biofilm: The Mode of Bacterial Life 118
5.1.1 Extracellular Polymeric Matrix 118
5.1.2 Extracellular Polysaccharide Matrix in Staphylococci 119
5.1.3 Extracellular Matrix in Pseudomonas Biofilm and Alginate 120
5.1.4 Extracellular Polysaccharides in Candida Biofilms 120
5.1.5 Production of Extracellular Polysaccharides in Streptococci, Lactobacilli, and Multispecies Oral Biofilms 121
5.2 Capillary Electrophoresis of Microbial Cells 122
5.2.1 The Surface Charge of Microorganisms 122
5.2.2 The Isoelectric Point of Microorganisms 124
5.2.3 Capillary Electromigration Techniques 124
5.2.3.1 Capillary Surface Modification 125
5.2.3.2 Detection of the Microorganisms in Capillary Electrophoresis 126
5.2.3.3 CZE of Microorganisms 126
5.2.3.4 CIEF of Microorganisms 128
5.3 Capillary Electromigration Techniques: A Useful Tool for the Detection of Biofilm Formation, an Important Virulence Factor in Microorganisms 128
References 132
Chapter 6: Capillary Electrophoresis Applied to Polysaccharide Characterization 139
6.1 Introduction 140
6.2 Structural Studies Via Degradation Mechanisms 141
6.3 Studies of Macromolecular Features 148
6.4 Capillary Electrophoresis for Practical Uses 153
6.5 Capillary Electrophoresis in Biomedical and Biological Application Fields 155
6.6 Conclusion 157
References 158
Chapter 7: Capillary Electrophoresis for Monitoring Natural and Synthetic Processes of Saccharide-Bearing Molecules 162
7.1 Introduction 163
7.2 Synthesis Involving Mono- and Oligo-saccharide Species 165
7.3 Glycoconjugate Preparation 170
7.3.1 Saccharidic Backbone 170
7.3.2 Non-saccharidic Backbone 172
7.4 Saccharidic Species as Enzyme Substrate: Studies on Enzyme Activity and Mechanism of Action 172
7.5 Studies of Natural Processes and of Reactions in Single Cells 177
7.6 Conclusion 179
References 179
Chapter 8: Analysis of Intact Glycoprotein Biopharmaceuticals by Capillary Electrophoresis 184
8.1 Introduction 185
8.2 General Considerations 187
8.3 Capillary Electrophoresis and Glycoprotein Pharmaceuticals 189
8.3.1 Impact of the Process 189
8.3.1.1 Host Cell or Expression System 189
8.3.1.2 Cell Culture Conditions 191
8.3.1.3 Purification Process 193
8.3.2 Quality Control 194
8.3.2.1 In-Process Monitoring 194
8.3.2.2 Purified Products 195
8.3.2.3 Finished Product 198
8.3.3 Stability Testing 204
8.4 CE in Biological Fluids for Clinical Application 206
8.5 Conclusion 209
References 210
Chapter 9: Capillary Electrophoresis and Capillary Electrophoresis–Mass Spectrometry for Structural Analysis of N-Glycans Derived from Glycoproteins 216
9.1 Introduction 218
9.2 Isolation and Direct Analysis of Intact Glycoproteins 220
9.3 Release of N-Glycans from Glycoproteins 223
9.3.1 Chemical Release 223
9.3.2 Enzymatic Release 224
9.4 Analysis of N-Glycans by Capillary Electrophoresis 224
9.4.1 Derivatization of Released N-Glycans and the Detection 224
9.4.2 Separation Modes 228
9.4.2.1 Capillary Zone Electrophoresis 229
9.4.2.2 Capillary Gel Electrophoresis 231
9.4.2.3 Capillary Isoelectric Focusing 231
9.4.2.4 Micellar Electrokinetic Chromatography 231
9.4.2.5 Capillary Electrochromatography 232
9.4.2.6 Capillary Affinity Electrophoresis 232
9.5 Analysis of N-Glycans by Capillary Electrophoresis–Mass Spectrometry 233
9.5.1 Interfacing with Mass Spectrometry 235
9.5.2 Derivatization of N-Glycans for Capillary Electrophoresis–Mass Spectrometry Analysis 236
9.5.3 Capillary Zone Electrophoresis–Mass Spectrometry 237
9.5.4 Capillary Electrochromatography–Mass Spectrometry 239
9.6 Conclusion 239
References 240
Chapter 10: Monosaccharide Compositional Analysisof Glycoproteins and Glycolipids: Utilityin the Diagnosis/Prognosis of Diseases 247
10.1 Introduction 248
10.2 Chemical and Enzymatic Release for Monosaccharide Compositional Analysis 249
10.3 Chromatographic and Electrophoretic Methods Developed for Monosaccharide Compositional Analysis of Glycoproteins and Glycolipids 251
10.3.1 High-Performance Anion Exchange Chromatography with Pulsed Amperometric Detection 251
10.3.2 High-Performance Liquid Chromatography with Fluorescence Detection 256
10.3.3 High-Performance Liquid Chromatography with Mass Spectrometry 262
10.3.4 Capillary Electrophoresis with Laser-Induced Fluorescence 265
10.4 Monosaccharide Compositional Analysis of Glycoproteins and Glycolipids in the Diagnosis/Prognosis of Diseases 268
10.5 Conclusion 270
References 271
Index 278