Methods in Molecular Biology

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 (R) Databases and Tools for Antibody Engineering and Humanization



Marie-Paule Lefranc, Francois Ehrenmann, Sofia Kossida, Veronique Giudicelli, and Patrice Duroux







Part II: Generation of Diversity







4. Construction of Human Naive Antibody Gene Libraries



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







5. Construction of Synthetic Antibody Libraries



Deborah 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 Koenning, Steffen Hinz, Julius Grzeschik, Christian Schroeter, 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 Pluckthun







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 Muller







Part V: Fc Engineering







19. Chemoenzymatic Defucosylation of Therapeutic Antibodies for Enhanced Effector Functions Using Bacterial α-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 Roesner, 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, Clement 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