Carbohydrate microarrays emerged as a key technology for the deciphering of the glycospace by providing a multiplex technology where tens to hundreds of carbohydrates/protein interactions can be probed in parallel. "Carbohydrate Microarrays: Methods and Protocols" aims to give the reader the theoretical and experimental clues necessary for the fabrication and implementation of carbohydrate microarrays. This requires three essential steps: to obtain the carbohydrate probes (monosacharides, oligosacchrides, polysacchairdes, glycoconjugates or glycoclusters); to immobilize these probes, and, to implement the protocols for biological/biochemical interaction with the desired target. This volume gives an overview of carbohydrate microarray and carbohydrate chemistry and illustrates different detection techniques and their applications.
Written in the successful "Methods in Molecular Biology" series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, "Carbohydrate Microarrays: Methods and Protocols" compiles a catalogue of protocols on carbohydrate microarrays to span the needs of researchers around the globe.
1. Recent Advances and Future Challenges in Glycan Microarray Technology
2. Chemical Synthesis of Carbohydrates and Their Surface Immobilisation: A Brief Introduction
3. General Consideration on Sialic Acid Chemistry
4. Synthesis of Azido-Functionalized Carbohydrates for the Design of Glycoconjugates
5. Polypyrrole-Oligosaccharide Microarray for the Measurement of Biomolecular Interactions by Surface Plasmon Resonance Imaging
6. Glycosylated Self-Assembled Monolayers for Arrays and Surface Analysis
7. Carbohydrate Microarrays for Enzymatic Reactions and Quantification of Binding Affinities for Glycan-Protein Interactions
8. Neoglycolipid (NGL)-Based Oligosaccharide Microarray System: Preparation of NGLs and Their Non-Covalent Immobilization on Nitrocellulose-Coated Glass Slides for Microarray Analyses
9. Preparation of a Mannose-6-phosphate Glycan Microarray Through Fluorescent Derivatization, Phosphorylation, and Immobilization of Natural High-mannose N-glycans and its Application in Ligand Identification of P-type Lectins 10. Production of Fluorous-Based Microarrays with Uncharged Carbohydrates
11. General Procedure for the Synthesis of Neoglycoproteins and Immobilization on Epoxide-Modified Glass Slides
12. Immobilization of Polyacrylamide-Based Glycoconjugates on Solid Phase in Immunosorbent Assays
13. Surface Plasmon Resonance Imaging Analysis of Protein Binding to a Sialoside-Based Carbohydrate Microarray
14. Glycoarray by DNA Directed Immobilization
15. Fabrication of Carbohydrate Surfaces by Using Non Derivatised Oligosaccharides
16. Polysaccharide Microarrays. Application to the Identification of Heparan Sulfate Mimetics
17. Carbohydrate Antigen Microarrays
18. Probing Virus-Glycan Interactions Using Glycan Microarrays
19. MALDI-ToF MS Analysis of Glycosyltransferase Activities on Gold Surface Arrays
20. Microarray Technology using Glycans Extracted from Natural Sources for Serum Antibody Fluorescent Detection
21. Studying Modification of Aminoglycoside Antibiotics by Resistance-causing Enzymes via Microarray
22. Microarray Method for the Rapid Detection of Glycosaminoglycan-Protein Interactions
23. Neoglycolipid-based 'Designer' Oligosaccharide Microarrays to Define b-glucan Ligands for Dectin-1
24. Measurement of Antibodies to Pneumococcal Polysaccharides with Luminex xMAP Microsphere-based Liquid Arrays
25. Carbohydrate Microarrays in 96-well Polystyrene Microtiter Plates
26. Photoimmobilization of Saccharides
27. Microwave Assisted Method for Fabrication of Carbohydrate Cluster Microarrays on 3-Dimensional Hydrazide-Dendrimer Substrate
28. Combinatorial Glycoarray