Includes cutting-edge techniques for the study of antibody engineering

Provides step-by-step detail essential for reproducible results

Contains updated key implementation advice from the experts

Part I: In Silico Methods

1. Antibody Design and Humanization via In Silico Modeling

Vinodh B. Kurella and Reddy Gali

2. Antibody Affinity Maturation by Computational Design

Daisuke Kuroda and Kouhei Tsumoto

3. Use of IMGT® Databases and Tools for Antibody Engineering and Humanization

Marie-Paule Lefranc, François Ehrenmann§, Sofia Kossida, Véronique Giudicelli, and Patrice Duroux

Part II: Generation of Diversity

4. Construction of Human Naïve Antibody Gene Libraries

Michela Pasello, Alessandra Mallano, Michela Flego, Silvia Zamboni, Anna Maria Giudice, and Katia Scotlandi

5. Construction of Synthetic Antibody Libraries

Déborah Caucheteur, Gautier Robin, Vincent Parez, and Pierre Martineau

6. Construction of Histidine-Enriched Shark IgNAR Variable Domain Antibody Libraries for the Isolation of pH-Sensitive vNAR Fragments

Doreen Könning, Steffen Hinz, Julius Grzeschik, Christian Schröter, Simon Krah, Stefan Zielonka, and Harald Kolmar

7. Display Technologies for Generation of Ig Single Variable Domains

Vladimir Bobkov, Bas van der Woning, and Hans de Haard

8. A Streamlined Approach for the Construction of Large Yeast Surface Display Fab Antibody Libraries

Simon Krah, Julius Grzeschik, Simon Rosowski, Ramona Gaa, Iris Willenbuecher, Deniz Demir, Lars Toleikis, Harald Kolmar, Stefan Becker, and Stefan Zielonka

Part III: Selections of Lead Candidate

9. Phage Display and Selections on Purified Antigens

Magali Colazet and Patrick Chames

10. Selection of Antibodies to Transiently Expressed Membrane Proteins Using Phage Display

Martina L. Jones, Stephen M. Mahler, and Sumukh Kumble

11. Selection of Antibody Fragments against Structured DNA by Phage Display

Mahdi Zeraati, Marcel E. Dinger, and Daniel Christ

12. Selection of Antibody Fragments by Yeast Display

Nathalie Scholler

13. Rapid Selection of High Affinity Antibody scFv Fragments Using Ribosome Display

Birgit Dreier and Andreas Plückthun

14. In Vitro Selection of Single Domain Antibody (VHH) Using cDNA Display

Naoto Nemoto, Shigefumi Kumachi, and Hidenao Arai

15. Sequencing and Affinity Determination of Antigen-Specific B Lymphocytes from Peripheral Blood

Peter Schofield, Rodrigo Vazquez-Lombardi, Mahmoud Abdelatti, Damien Nevoltris, Christopher C. Goodnow, Daniel Christ, and Joanne H. Reed

Part IV: Production of Recombinant Antibodies

16. Expression of IgG Monoclonals with Engineered Immune Effector Functions

Rodrigo Vazquez-Lombardi, Damien Nevoltris, Romain Rouet, and Daniel Christ

17. An IRES-Mediated Tricistronic Vector for Efficient Generation of Stable, High Level Monoclonal Antibody Producing CHO DG44 Cell Lines

Jessna H.M. Yeo, Mariati, and Yuansheng Yang

18. Production, Purification, and Characterization of Antibody-TNF Superfamily Ligand Fusion Proteins

Martin Siegemund, Nadine Beha, and Dafne Müller

Part V: Fc Engineering

19. Chemoenzymatic Defucosylation of Therapeutic Antibodies for Enhanced Effector Functions Using Bacterial ¿-Fucosidases

Chao Li, Tiezheng Li, and Lai-Xi Wang

20. Fc Glyco- and Fc Protein-Engineering: Design of Antibody Variants with Improved ADCC and CDC Activity

Christian Kellner, Stefanie Derer, Katja Klausz, Sophia Rosskopf, Tim Wirt, Thies Rösner, Anna Otte, Elisa Cappuzzello, and Matthias Peipp

21. Fc Engineering: Tailored Synthetic Human IgG1-Fc Repertoire for High Affinity Interaction with FcRn at pH 6.0

Abhishek Saxena, Bingxin Bai, Shin-Chen Hou, Lianlian Jiang, Tianlei Ying, Shane Miersch, Sachdev S. Sidhu, and Donghui Wu

Part VI: Characterization, Optimization, and Innovative Formats

22. Measuring Antibody-Antigen Binding Kinetics Using Surface Plasmon Resonance

Stephen Hearty, Paul Leonard, Hui Ma, and Richard O¿Kennedy

23. Parallel Evolution of Antibody Affinity and Thermal Stability for Optimal Biotherapeutic Development

Edward Franklin, Orla Cunningham, and Brian Fennell

24. The Use of Somatic Hypermutation for the Affinity Maturation of Therapeutic Antibodies

Peter M. Bowers, William J. Boyle, and Robert Damoiseaux

25. Selection and Use of Intracellular Antibodies

Sandrine Moutel, Clément Nizak, and Franck Perez

26. Site-Specific Radioactive Labeling of Nanobodies

Maxine Crauwels, Sam Massa, Charlotte Martin, Cecilia Betti, Steven Ballet, Nick Devoogdt, Catarina Xavier, and Serge Muyldermans