Description
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 Biology(TM) 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. 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 Ma, Sandrine Donadio-Andri, Chlo Iss, Valrie 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, Marylne Lvque, Pierre Crutti, Annick Ozil, Sylvie Choblet, Marie-Luce Violet, Marie-Christine Slomianny, Anne Harduin-Lepers, and Martine Crutti 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 Corvaa, 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, Cline Huillet, Olivier Colas, Christine Klinguer-Hamour, Nathalie Corvaa, 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 Gry 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 Sjblom, 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 Cdric Atmanene, Elsa Wagner-Rousset, Nathalie Corvaa, 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 Franois Becher, Mathieu Dubois, Franois Fenaille, and Eric Ezan
