ISBN-13: 9781493959440 / Angielski / Miękka / 2016 / 734 str.
ISBN-13: 9781493959440 / Angielski / Miękka / 2016 / 734 str.
Recent medical advances have opened up exciting new possibilities for antibodies as a key drug modality in diseases including cancer and neurodegenerative disorders. This updated second edition has detailed expert guidance on the latest methods and protocols.
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
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.
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