High-Throughput Glycomics and Glycoproteomics, 1st ed. 2017
Methods and Protocols
Methods in Molecular Biology Series, Vol. 1503

This volume discusses detailed protocols for the analysis of glycosylation at the level of free glycans and glycopeptides. The book covers topics such as the importance of glycans in eukaryotic life (with a focus on mammals and particularly humans); recent technologies that allow the characterization of larger sets of samples; and method repeatability and robustness, as well as higher throughout with respect to sample preparation, measurement and data analysis. The chapters spans a wide range of techniques, including the analysis of fluorescently labeled glycans with HPLC, LC-MS analysis of glycopeptides both for glycosylation profiling and for in-depth tandem mass spectrometric analysis of protein glycosylation, and the analysis of glycans with fluorescent labeling, capillary electrophoresis in conjunction with laser induced fluorescence detection. The chapters also detail specific samples types including brain tissues, N-glycans from in-vitro cell cultures, milk oligosaccharides, invertebrate and protest N-glycans, as well as plant glycans. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction 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.

Practical and thorough, High-Throughout Glycomics and Glycoproteomics: Methods and Protocols is an essential reference for researchers planning to enter this rapidly evolving field.

1. Ubiquitous Importance of Protein Glycosylation
Jasminka Krištić and Gordan Lauc

2. Laboratory Experimental Design for a Glycomic Study
Ivo Ugrina, Harry Campbell,, and Frano Vučković

3. High-Throughput Analysis of the IgG N-Glycome by UPLC-FLR
Maja Pučić-Baković

4. High-Throughput Analysis of IgG Fc Glycopeptides by LC-MS
David Falck, Bas C. Jansen, Noortje de Haan, and Manfred Wuhrer

5. Sialic Acid Derivatization for the Rapid Subclass and Sialic Acid Linkage-Specific MALDI-TOF-MS Analysis of IgG Fc-Glycopeptides
Noortje de Haan, Karli R. Reiding, and Manfred Wuhrer

6. The Use of Multiple Reaction Monitoring on QQQ-MS for the Analysis of Protein- and Site-Specific Glycosylation Patterns in Serum
L. Renee Ruhaak

7. Analysis of Permethylated Glycan by Liquid Chromatography (LC) and Mass Spectrometry (MS)
Shiyue Zhou, Kerry Wooding, and Yehia Mechref

8. High-Throughput Analysis of the Plasma N-Glycome by UHPLC
Barbara Adamczyk, Henning Stöckmann, Róisín O’Flaherty, Niclas G. Karlsson, and Pauline M. Rudd

9. Site-Specific N- and O-Glycopeptide Analysis Using an Integrated C18-PGC-LC-ESI-QTOF-MS/MS Approach
Kathrin Stavenhagen, Hannes Hinnburg, Daniel Kolarich, and Manfred Wuhrer

10. Analysis of Milk Oligosaccharides by Mass Spectrometry
Lauren D. Wu, L. Renee Ruhaak, and Carlito B. Lebrilla

11. N- and O-Glycomics from Minor Amounts of Formalin-Fixed, Paraffin-Embedded Tissue Samples
Hannes Hinneburg, Falko Schirmeister, Petra Korać, and Daniel Kolarich

12. Carbohydrate Microarray Technology Applied to High-Throughput Mapping of Plant Cell Wall Glycans Using Comprehensive Microarray Polymer Profiling (CoMPP)
Stjepan Krešimir Kračun, Jonatan Ulrik Fangel, Maja Gro Rydahl, Henriette Lodberg Pedersen, Silvia Vidal-Melgosa, and William George Tycho Willats

13. Analysis of Invertebrate and Protist N-Glycans
Alba Hykollari, Katharina Paschinger, Barbara Eckmair, and Iain B.H. Wilson

14. High-Throughput and High-Sensitivity Mass Spectrometry-Based N-Glycomics of Mammalian Cells
Stephanie Holst, Gabi W. Van Pelt, Wilma E. Mesker, Rob A. Tollenaar, Ana I. Belo, Irma van Die, Yoanna Rombouts, and Manfred Wuhrer

15. Analysis of N-Glycosylation of Total Membrane Proteins
Tamara Pavić and Olga Gornik

16. HILIC-UPLC Analysis of Brain Tissue N-Glycans
Thomas Klarić and Ivan Gudelj

17. Automated Integration of a UPLC Glycomic Profile
Anna Agakova, Frano Vuckovic, Lucija Klaric, Gordan Lauc, and Felix Agakov

18. Databases and Associated Tools for Glycomics and Glycoproteomics
Frederique Lisacek, Julien Mariethoz, Davide Alocci, Pauline M. Rudd, Jodie L. Abrahams, Matthew P. Campbell, Nicolle H. Packer, Jonas Ståhle, Göran Widmalm, Elaine Mullen, Barbara Adamczyk, Miguel Rojas Macias, Chunsheng Jin, and Miclas G. Karlsson

19. High-Throughput N-Glycan Analysis with Rapid Magnetic Bead-Based Sample Preparation
Marton Szigeti and Andras Guttman

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts

Includes supplementary material: sn.pub/extras