Antibody Engineering. Methods and Protocols (Methods in Molecular Biology, Vol. 907)

Description

• Includes cutting-edge methods and protocols
• Provides step-by-step detail essential for reproducible results
• Contains key notes and implementation advice from the experts

More than ever, antibodies are being recognized as a major drug modality in a variety of diseases, including cancer, autoimmune diseases, infectious diseases, or even neurodegenerative disorders. Over 30 therapeutic antibodies have been approved and novel molecules are entering clinical trials at an average rate of 50 per year and that is predicted to continue well into the future. Notwithstanding the many achievements already made in the field, there is still a lot of room for improvements for these molecules in terms of activity, and a plethora of approaches have been attempted to optimize these molecules. Antibody Engineering: Methods and Protocols, Second Edition was compiled to give complete and easy access to a variety of antibody engineering techniques, starting from the creation of antibody repertoires and efficient ways to select binders from these repertoires, to their production in various hosts, their detailed characterization using various well established techniques, and to the modification and optimization of these lead molecules in terms of binding activity, specificity, size, shape, and more. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, Antibody Engineering: Methods and Protocols, Second Edition serves as an invaluable resource for both experts and those new to the field, and most of all as a source of inspiration for the creation of the antibodies of tomorrow.

Content Level» Professional/practitioner
Keywords» Fc engineering - antibody repertoires - antibody variable domains - in silico methods - recombinant antibodies - therapeutic antibodies
Related subjects » Immunology

TABLE OF CONTENTS

Part I. In silico Methods

• 1. Use of IMGT® Databases and Tools for Antibody Engineering and Humanization
  Marie-Paule Lefranc, François Ehrenmann, Chantal Ginestoux, Véronique Giudicelli, and Patrice Duroux
• 2. Computer-Assisted Modeling of Antibody Variable Domains
  Oscar H. P. Ramos

Part II. Generation of Diversity

• 3. Cloning Single-Chain Antibody Fragments (scFv) from Hybridoma Cells
  Lars Toleikis and André Frenzel
• 4. Human in Cell scFv Library from Infiltrating B Cell
  Sylvie Peraldi-Roux
• 5. Construction of Human Naive Antibody Gene Libraries
  Michael Hust, André Frenzel, Torsten Meyer, Thomas Schirrmann, and Stefan Dübel
• 6. Synthetic Customized scFv Libraries
  Gautier Robin and Pierre Martineau
• 7. Selection of Stable scFv Antibodies by Phage Display
  Eeva-Christine Brockmann
• 8. Generation of Single Domain Antibody Fragments Derived from Camelids and Generation of Manifold Constructs
  Cécile Vincke, Carlos Gutiérez, Ulrich Wernery, Nick Devoogdt, Gholamreza Hassanzadeh-Ghassabeh, and Serge Muyldermans
• 9. Generation and Isolation of Target Specific Single Domain Antibodies from Shark Immune Repertoires
  Mischa Roland Müller, Ronan O’Dwyer, Marina Kovaleva, Fiona Rudkin, Helen Dooley, and Caroline Jane Barelle
• 10. Generation of Human Single Domain Antibody Repertoires by Kunkel Mutagenesis
  Kip Dudgeon, Romain Rouet, Kristoffer Famm, and Daniel Christ

Part IV. Selections of Lead Candidate

• 11. Phage Display and Selections on Purified Antigens
  Julie Matz and Patrick Chames
• 12. Phage Display and Selections on Cells
  Klervi Even-Desrumeaux and Patrick Chames
• 13. Humanization by CDR Grafting and Specificity-Determining Residue Grafting
  Jin Hong Kim and Hyo Jeong Hong
• 14. Humanization by Guided Selections
  Sang Jick Kim and Hyo Jeong Hong
• 15. Selection of Antibody Fragments by Yeast Display
  Nathalie Scholler
• 16. Evolution of Antibodies in vitro by Ribosome Display
  Bryan M. Edwards and Mingyue He
• 17. Mammalian Cell Display of Full Length IgG
  Chen Zhou and Wenyan (David) Shen

Part V. Production of Recombinant Antibodies

• 18. Production of Antibody Fragments in Escherichia coli
  Tomohisa Katsuda, Hiroyuki Sonoda, and Hideki Yamaji
• 19. Production of Antibody Derivatives in the Methylotrophic Yeast Pichia pastoris
  Steve Schoonooghe, Jannick Leoen, and Jurgen Haustraete
• 20. Monoclonal Antibody Expression in Mammalian Cells
  Richard (Yi) Zhang and Wenyan (David) Shen
• 21. Production of Recombinant Antibodies in Drosophila melanogaster S2 Cells
  Daniel X. Johansson, Thomas Krey, and Oskar Andersson
• 22. Production of Antibody Fragments Using the Baculovirus-Insect Cell System
  Takanori Furuta, Takafumi Ogawa, and Hideki Yamaji
• 23. Transient and Stable Expression of Antibodies in Nicotiana Species
  J. Christopher Hall, Michael D. McLean and Freydoun Garabagi

Part VI. Variable Domain Optimization

• 24. Measuring Antibody-Antigen Binding Kinetics Using Surface Plasmon Resonance
  Stephen Hearty, Paul Leonard, and Richard O’Kennedy
• 25. Affinity Determination of Biotinylated Antibodies by Flow Cytometry
  Klervi Even-Desrumeaux and Patrick Chames
• 26. Affinity Maturation of Antibodies: Optimized Methods to Generate High Quality scFv Libraries and Isolate IgG Candidates by High Throughput Screening
  Laurence Renaut, Céline Monnet, Olivier Dubreuil, Ouafa Zaki, Fabien Crozet, Khalil Bouayadi, Hakim Kharrat and Philippe Mondon
• 27. Affinity Maturation by Semi Rational Approaches
  Rodrigo Barderas, Johan Desmet, Philippe Alard, and J. Ignacio Casal
• 28. Molecular Scanning Mutagenesis: Combining Ribosome Display, Random Mutagenesis and Bioinformatic Analysis for Protein Engineering
  Alfredo Darmanin-Sheehan, William James Jonathan Finlay Orla Cunningham, and Brian Joseph Fennell

Part VII. Fc Engineering

• 29. Fucose-Targeted Glyco-Engineering of Pharmaceutical Cell Lines
  Christiane Ogorek, Ingo Jordan, Volker Sandig, and Hans Henning von Horsten
• 30. Fc engineering: Design, expression and functional characterization of antibody variants with improved effector function
  Stefanie Derer, Christian Kellner, Sven Berger, Thomas Valerius and Matthias Peipp
• 31. Fc engineering: Serum half life modulation through FcRn binding
  Tove Olafsen
• 32. Monoclonal Antibody Lead Characterization - In vitro and in vivo methods
  Axel Hernandez, Julie Parmentier, Youzhen Wang, Jane Cheng and Gadi Gazit Bornstein

Part VIII. Innovative Formats

• 33. Production, generation and characterization of scFv TNF ligand fusion proteins
  Andrea Fick, Agnes Wyzgol and Harald Wajant
• 34. Antibody-IL-2 fusion proteins for tumor targeting
  Andreas A. Hombach and Hinrich Abken
• 35. Recombinant immunotoxins containing low endotoxins for clinical and animal studies
  Masanori Onda
• 36. Chimeric antigen receptors for T-cell based therapy
  Eleanor J. Cheadle, Vicky Sheard, Andreas A. Hombach, Markus Chmielewski, Tobias Riet, Cor Berrevoets, Erik Schooten, Cor Lamers, Hinrich Abken, Reno Debets, David E. Gilham
• 37. Selection and Use of Intracellular Antibodies (Intrabodies)
  Sandrine Moutel, Clément Nizak, and Franck Perez
• 38. Radiolabeled Antibodies for Imaging and Therapy
  Jacques Barbet, Manuel Bardiès, Mickael Bourgeois, Jean-François Chatal, Michel Chérel, François Davodeau, Alain Faivre-Chauvet, Jean-François Gestin, and Françoise Kraeber-Bodéré
• 39. Design and Production of Multimeric Antibody Fragments, Focused on Diabodies with Enhanced Clinical Efficacy
  Glenn A. Powers, Peter J. Hudson, and Michael P. Wheatcroft
• 40. Production of Bispecific Antibodies: Diabodies and Tandem scFv
  Nora Hornig and Aline Färber-Schwarz