Glycosylation Engineering of Biopharmaceuticals: Methods and Protocols by Alain BeckGlycosylation Engineering of Biopharmaceuticals: Methods and Protocols by Alain Beck

Glycosylation Engineering of Biopharmaceuticals: Methods and Protocols

byAlain Beck

Hardcover | March 9, 2013

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Glyco-engineering is being developed as a method to control the composition of carbohydrates and to enhance the pharmacological properties of monoclonal antibodies (mAbs) and other proteins.  In Glycosylation Engineering of Biopharmaceuticals: Methods and Protocols, experts in the field provide readers with production and characterization protocols of glycoproteins and glyco-engineered biopharmaceuticals with a focus on mAbs. The volume is divided in four complementary parts dealing with glyco-engineering of therapeutic proteins, glycoanalytics, glycoprotein complexes characterization, and PK/PD assays for therapeutic antibodies.  Written in the highly successful Methods in Molecular BiologyT 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.


Authoritative and cutting-edge, Glycosylation Engineering of Biopharmaceuticals: Methods and Protocols serves as an ideal guide for scientists striving to push forward the exciting field of engineered biopharmaceuticals.
Title:Glycosylation Engineering of Biopharmaceuticals: Methods and ProtocolsFormat:HardcoverDimensions:355 pagesPublished:March 9, 2013Publisher:Springer-Verlag/Sci-Tech/TradeLanguage:English

The following ISBNs are associated with this title:

ISBN - 10:1627033262

ISBN - 13:9781627033268

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Table of Contents

Part I: Glyco-Engineering of Therapeutic Proteins


1. Engineering of Therapeutic and Diagnostic O-Glycans on Recombinant Mucin-Type Immunoglobulin Fusion Proteins Expressed in CHO Cells

            Linda Lindberg, Jining Liu, and Jan Holgersson


2. Engineering a Human-Like Glycosylation to Produce Therapeutic Glycoproteins Based on 6-Linked Sialylation in CHO Cells

            Nassimal El Maï, Sandrine Donadio-Andréi, Chloé Iss, Valérie Calabro, and Catherine Ronin


3. Glycoengineered Pichia-Based Expression of Monoclonal Antibodies

            Dongxing Zha


4. N-Glycosylation Humanization for Production of Therapeutic Recombinant Glycoproteins in Saccharomyces cerevisiae

            Christelle Arico, Christine Bonnet, and Christophe Javaud


5. Engineering the Baculovirus Genome to Produce Galactosylated Antibodies in Lepidopteran Cells

            Sylvie Juliant, Marylêne Lévêque, Pierre Cérutti, Annick Ozil, Sylvie Choblet, Marie-Luce Violet, Marie-Christine Slomianny, Anne Harduin-Lepers, and Martine Cérutti


Part II: Glycoanalytics


6. Nano LC Chips MS/MS for the Characterization of N-Glycopeptides Generated from Trypsin Digestion of a Monoclonal Antibody

            Elsa Wagner-Rousset, Christine Schaeffer-Reiss, Audrey Bednarczyk, Nathalie Corvaïa, Alain Van Dorsselaer, and Alain Beck


7. Cetuximab Fab and Fc N-Glycan Fast Characterization Using IdeS Digestion and Liquid Chromatography Coupled to Electrospray Ionization Mass Spectrometry

            Marie-Claire Janin-Bussat, Laure Tonini, Céline Huillet, Olivier Colas, Christine Klinguer-Hamour, Nathalie Corvaïa, and Alain Beck


8. Therapeutic Antibody Glycosylation Analysis: A Contract Research Organization Perspective in the Frame of Batch Release or Comparability Support

            Arnaud Delobel, Fabrice Cantais, Anicet Catrain, Erell Dereux, and Géry Van Vyncht


9. Mass Spectrometric Analysis of O-Linked Oligosaccharides from Various Recombinant Expression Systems

            Diarmuid T. Kenny, Stefan Gaunit, Catherine A. Hayes, Anki Gustafsson, Magnus Sjöblom, Jan Holgersson, and Niclas G. Karlsson


10. Assessing Fc Glycan Heterogeneity of Therapeutic Recombinant Monoclonal Antibodies Using NP-HPLC

            T. Shantha Raju


11. Application of Capillary Electrophoresis in Glycoprotein Analysis

            Richard R. Rustandi, Carrie Anderson, and Melissa Hamm


12. Characterization of Glycoprotein Biopharmaceutical Products by Caliper LC90 CE-SDS Gel Technology

            Grace Chen, Sha Ha, and Richard R. Rustandi


13. Hydrophobic Interaction Chromatography to Analyze Glycoproteins

            Richard R. Rustandi


14. Lectin Glycoprofiling of Recombinant Therapeutic Interleukin-7

            Ludovic Landemarre and Eric Duverger


15. Analysis of Monoclonal Antibodies by Sedimentation Velocity Analytical Ultracentrifugation

            W. Blaine Stine, Jr.


Part III: Glycoprotein Complexes Characterization


16. Noncovalent Mass Spectrometry for the Characterization of Antibody/Antigen Complexes

            Cédric Atmanene, Elsa Wagner-Rousset, Nathalie Corvaïa, Alain Van Dorsselaer, Alain Beck, and Sarah Sanglier-Cianferani


17. Conformational Analysis of Recombinant Monoclonal Antibodies with Hydrogen/Deuterium Exchange Mass Spectrometry

            Damian Houde and John R. Engen


18. Epitope Mapping of Antibodies by Mass Spectroscopy: A Case Study

            Victor H. Obungu, Valentina Gelfanova, and Lihua Huang


Part IV: PK/PD Assays for Therapeutic Antibodies


19. Evaluation of Antibody-Dependent Cell Cytotoxicity Using Lactate Deshydrogenase (LDH) Measurement

            Matthieu Broussas, Lucile Broyer, and Liliane Goetsch


20. Evaluation of Complement Dependent Cytotoxicity Using ATP Measurement and C1q / C4b Binding

            Lucile Broyer, Liliane Goetsch, and Mattthieu Broussas


21. Capture of the Human IgG1 Antibodies by Protein A for the Kinetic Study of h-IgG / FcgR Interaction Using SPR-Based Biosensor Technology

            Thierry Champion and Alain Beck


22. Mass Spectrometry Protocol for the Absolute Quantification of a Monoclonal Antibody in Serum with Immunopurification

            François Becher, Mathieu Dubois, François Fenaille, and Eric Ezan

Editorial Reviews

From the reviews:"The book is intended as a reference manual of protocols and best practices for the practicing bench scientist. The authors do a marvelous job of covering a wide variety of tools and techniques and bringing them together in one volume. . This collection of protocols will find an audience with experimentalists in the biopharmaceutical industry as well as various academic labs. . Overall, I would strongly recommend this book . ." (Bibhash Mukhopadhyay, Frontiers in Pharmacology, September, 2013)"This book . useful as it details in one place the current state of art technologies and techniques for antibodies. I enjoyed reading the introductions to each chapter, which were accessible and gave an historical setting and overall aim for the methodologies that are set out in the chapter. . this book offers a good reference guide for those of us working in this area." (Sian Estdale, BTS Newsletter - The British Toxicology Society, Issue 43, Winter, 2013)