Glycosylation Methods and Protocols

This volume explores the latest techniques and methods used for performing up-to-date glycosylation research. The chapters in this book are organized into four parts. Part One looks at the latest analytical and bioinformatics technologies that enable the characterization of glycosylation complexity. Part Two details the importance of synthetic chemistry and glycoengineering in the fields of bioprocessing and biotherapeutic development. Part Three discusses systems biology and computational technologies used by scientists to analyze glycosylation events in the cell. Part Four focuses on how cellular glycosylation biomarkers can be identified and used to characterize human clinical datasets. Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Cutting-edge and practical, Glycosylation: Methods and Protocols is a valuable resource for any scientist or researcher interested in learning more about this exciting and developing field. 

Preface…

Table of Contents…

Contributing Authors…

Part I Analytical and Bioinformatics Methods for Studying Glycosylation

1. Glycoinformatics Tools for Comprehensive Characterization of Glycans Enzymatically Released from Proteins

2. GlycoStore: A Platform for H/UPLC and Capillary Electrophoresis Glycan Data 

Matthew P. Campbell, Sophie Zhao, Jodie L. Abrahams, Terry Nguyen-Khuong, and Pauline M. Rudd

3. An Interactive View of Glycosylation 

Julien Mariethoz, Davide Alocci, Niclas G. Karlsson, Nicolle H Packer, and Frédérique Lisacek

4. Characterization and Analysis of Food-Sourced Carbohydrates

Leonie J. Kiely and Rita M. Hickey

5. Comparative Glycomics Analysis of Mass Spectrometry Data

Yusen Zhou and Sriram Neelamegham 

Part II Synthetic Biology and Glycoengineering Methods for Controlling Glycosylation 

6. Cell Free Remodeling of Glycosylation of Antibodies

Letícia Martins Mota, Venkata Tayi, and Michael Butler

7. Rapid Antibody Glycoengineering in CHO Cells via RNA Interference and CGE-LIF N-Glycomics

Pavlos Kotidis, Masue Marbiah, Roberto Donini, Itzcóatl A. Gómez, Ioscani Jimenez del Val, Stuart M. Haslam, Karen M. Polizzi, and Cleo Kontoravdi

8. In Silico Analysis of Therapeutic Antibody Aggregation and the Influence of Glycosylation

Hyesoo Jeon, Jerrard M. Hayes, and  K. H. Mok 

Anjali Shenoy and Adam W. Barb

Part III Systems Biology and Computational Methods for Deciphering Glycosylation Networks

10. Computational Modeling of Glycan Processing in the Golgi for Investigating Changes in the Arrangements of Biosynthetic Enzymes

11. O-Glycologue: A Formal-Language-Based Generator of O-Glycosylation Networks

Andrew G. McDonald and Gavin P. Davey

12. Synthetic Strategies for FRET-Enabled Carbohydrate Active Enzyme Probes

Meenakshi Singh, Michael Watkinson, Eoin M. Scanlan, and Gavin J. Miller

Part IV Glycosylation Biomarkers and Clinical Datasets 

13. N-Glycan Characterization by Liquid Chromatography Coupled with Fluorimetry and Mass Spectrometry

Richard A. Gardner, Paulina A. Urbanowicz, and Daniel I. R. Spencer

14. Lectin Histochemistry for Tissues and Cells, and Dual Lectin and Antibody Co-Localization

Siobhán S. McMahon and Michelle Kilcoyne

15. Lectin Affinity Chromatography for the Discovery of Novel Cancer Glycobiomarkers: A Case Study with PSA Glycoforms and Prostate Cancer

Esther Llop and Rosa Peracaula

16. Metabolic Labelling of Primary Neurons using Carbohydrate Click-Chemistry

Jerrard M. Hayes, Darren M. O’Hara, and Gavin P. Davey

17. Monitoring the Sialome on Human Immune Cells

Laura K. O’Farrell, Alexander Fraser, and Gavin P. Davey

Subject Index List...