Mass Spectrometry of Glycoproteins

1. Front Matter

   Pages i-xiv

   PDF

2. Glycosylation of Therapeutic Proteins: A Critical Quality Attribute

   • Arnaud Delobel

   Pages 1-21

3. Characterization of Protein Glycoforms at Intact Level by Orbitrap Mass Spectrometry

   • Dan Bach Kristensen, Trine Meiborg Sloth, Martin Ørgaard, Pernille Foged Jensen

   Pages 23-45

4. Analysis of Intact Glycoproteins by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

   • Estela Giménez, Montserrat Mancera-Arteu, Fernando Benavente, Victoria Sanz-Nebot

   Pages 47-56

5. Fc Glycosylation Characterization of Human Immunoglobulins G Using Immunocapture and LC-MS

   • Yosra Helali, Shilpee Sharma, Marie Vandeput, Dansala Welba, Pierre Van Antwerpen, Arnaud Marchant et al.

   Pages 57-71

6. Fast Afucosylation Profiling of Glycoengineered Antibody Subunits by Middle-Up Mass Spectrometry

   • Elsa Wagner-Rousset, Olivier Colas, Stéphane Chenu, Yannis-Nicolas François, Davy Guillarme, Sarah Cianferani et al.

   Pages 73-83

7. Characterization of Glycosylated Proteins at Subunit Level by HILIC/MS

   • Valentina D’Atri, Davy Guillarme

   Pages 85-95

8. Analysis of Monoclonal Antibody Glycopeptides by Capillary Electrophoresis–Mass Spectrometry Coupling (CE-MS)

   • Josiane Saadé, Michael Biacchi, Jérémie Giorgetti, Antony Lechner, Alain Beck, Emmanuelle Leize-Wagner et al.

   Pages 97-106

9. Enrichment of Intact Glycopeptides Using Strong Anion Exchange and Electrostatic Repulsion Hydrophilic Interaction Chromatography

   • Abel Bermudez, Sharon J. Pitteri

   Pages 107-120

10. In-Depth Glycan Characterization of Therapeutic Glycoproteins by Stepwise PGC SPE and LC-MS/MS

    • Myung Jin Oh, Youngsuk Seo, Unyong Kim, Hyun Joo An

    Pages 121-131

11. Site-Specific N-glycosylation Analysis of Recombinant Proteins by LC/MS^E

    • Kevin Canis, Estelle Garénaux, Jean-François Boe

    Pages 133-154

12. Mapping O-glycosylation Sites Using OpeRATOR and LC-MS

    • Maria Nordgren, Andreas Nägeli, Helén Nyhlén, Jonathan Sjögren

    Pages 155-167

13. Site-Specific O-Glycosylation Analysis by Liquid Chromatography–Mass Spectrometry with Electron-Transfer/Higher-Energy Collisional Dissociation

    • Noritaka Hashii, Junya Suzuki

    Pages 169-178

14. Profiling of N-Linked Oligosaccharides of a Glycoprotein by UPLC-FLR-ESI-MS After Derivatization with Fluorescent Anthranilamide

    • Claire I. Butré, Eric Largy, Arnaud Delobel

    Pages 179-188

15. Evaluating N-Glycosylation of a Therapeutic Monoclonal Antibody Using UHPLC-FLR-MS with RapiFluor-MS Labeling

    • Rosie Upton, James Duffy, Sam Clawson, David Firth

    Pages 189-203

16. Routine Analysis of N-Glycans Using Liquid Chromatography Coupled to Routine Mass Detection

    • Ximo Zhang, Vithiya Vimalraj, Mitul Patel

    Pages 205-219

17. Online 2D-LC for Complex N-Glycan Analysis from Biopharmaceuticals

    • Sonja Schneider, Edgar Naegele, Sonja Krieger

    Pages 221-235

18. Profiling, Relative Quantification, and Identification of Sialylated N-Linked Oligosaccharides by UPLC-FLR-ESI/MS After Derivatization with Fluorescent Anthranilamide

    • Claire I. Butré, Eric Largy, Fabrice Cantais, Arnaud Delobel

    Pages 237-247

19. Linkage Analysis of Oligosaccharides and Polysaccharides: A Tutorial

    • Ian Black, Christian Heiss, Russell W. Carlson, Parastoo Azadi

    Pages 249-271

20. Use of Exoglycosidases for the Structural Characterization of Glycans

    • Elizabeth McLeod, Paula Magnelli, Xiaofeng Shi

    Pages 273-280

This volume presents methods used for the analysis of glycoproteins at different levels—intact, subunit, glycopeptide, and monosaccharide--, and discusses and solves most analytical challenges that a scientist working on glycoproteins may come across. The chapters in this book cover topics such as the role of glycosylation on the properties of therapeutic glycoproteins; different analytical methods to characterize glycosylation, from the intact proteins to the glycan level, for both N-linked and O-linked glycoproteins; mass spectrometry imaging methodology for glycosylation analysis in tissues; approaches to characterizing glycosylation on cultured cells; and the use of cloud computing to deploy mass spectrometry data analysis. 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 thorough, Mass Spectrometry of Glycoproteins: Methods and Protocols is a valuable resource for scientists interested in learning more about this developing field.

• glycosylation
• HILIC-MS
• Bioinformatics
• MALDI-MS
• N-glycans

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  Arnaud Delobel

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