Human Monoclonal Antibodies (2^nd Ed., 2nd ed. 2019) Methods and Protocols Methods in Molecular Biology Series, Vol. 1904

This second edition volume expands on the previous edition with descriptions of recent developments in the field. The chapters in this book cover topics such as monoclonal antibodies for the treatment of melanoma; production and purification of human monoclonal antibodies; humanization and optimization of monoclonal antibodies; rapid chimerization of monoclonal antibodies; epitope mapping via phage display from single gene libraries; recombinant antibodies made by combining phage and yeast display selections; production of stabilized antibody fragments in the E. coli bacterial cytoplasm and transfected mammalian cells; and analysis of CAR T cells. Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Unique and thorough, Human Monoclonal Antibodies: Methods and Protocols, Second Edition is a valuable tool for novice and expert researchers interested in learning more about this evolving field.

Dedication…

Preface…

Table of Contents…

Contributing Authors…

1. Human Monocolonal Antibodies: The Benefits of Humanization

Herman Waldmann

2. Cancer Immunotherapy: The Dawn of Antibody Cocktails

Ilaria Marrocco, Donatella Romaniello, and Yosef Yarden

3. IgM Natural Autoantibodies in Physiology and the Treatment of Disease

Mahboobeh Fereidan-Esfahani, Tarek Nayfeh, Arthur Warrington, Charles L. Howe, and Moses Rodriguez

4. Monoclonal Antibodies for the Treatment of Melanoma: Present and Future Strategies

Madhuri Bhandaru and Anand Rotte

5. An Efficient Method to generate Monoclonal Antibodies from Human B Cells

Jenna J. Guthmiller, Haley L. Dugan, Karlynn E. Neu, Linda Yu-Ling Lan, and Patrick C. Wilson

6. Isolation of Antigen-Specific, antibody-Secreting Cells using a Chip-Based Immunospot Array

Hiroyuki Kishi, Tatsuhiko Ozawa, Hiroshi Hamana, Eiji Kobayashi, and Atsushi Muraguchi

7. Purification of Human Monoclonal antibodies and Their Fragments

Nicole Ulmer, Sebastian Vigg, Thomas Müller-Späth, and Massimo Morbidelli

8. One-Tube Multicolor Flow Cytometry Assay (OTMA) for Comprehensive Immunophenotyping of Peripheral Blood

Anna-Jasmina Donaubauer, Paul F. Rühle, Ina Becker, Rainer Fietkau, Udo S. Gaipl, and Benjamin Frey

9. Humanization and Simultaneous Optimization of Monoclonal Antibody

T. Kuramochi, T. Igawa, H. Tsunoda, and K. Hattori

10. Antibody Fragments Humanization: Beginning with the End in Mind

Nicolas Aubrey and Philippe Billiald

11. Antigen-Specific Human Monoclonal Antibodies from Transgenic Mice

Susana Magadán Mompó and África González-Fernández

12. Refining the Quality of Monoclonal Antibodies: Grafting Unique Peptide-Binding  Site in the Fab Framework

Jeremy D. King and John C. Williams

13. Basic Procedures for Detection and Cytotoxicity of Chimeric Antigen Receptors

Keichiro Mihara, Tetsumi Yoshida, and Joyeeta Bhattacharyya

14. Rapid Chimerization of Antibodies

Koji Hashimoto, Kohei Kurosawa, Hidetaka Seo, and Kunihiro Ohta

15. Phage Display Technology for Human Monoclonal Antibodies

Dal Ferro Marco, Rizzo Serena, Rizzo Emanuela, Marano Francesca, Luisis Immacoloata, Tarasiuk Olga, and Daniele Sblattero

16. Recombinant Antibody Selections by Combining Phage and Yeast Display

Fortunato Ferrara, Maria Felicia Soluri, and Daniele Sblattero

17. Epitope Mapping via Phage Display from Single Gene Libraries

Viola Fühner, Philip Alexander Heine, Kilian Zilkens, Doris Meier, Kristian Daniel Ralph Roth, Gustavo Marçal Schmidt Garcia Moreira, Michael Hust, and Giulio Russo

18. A High-Throughput Magnetic Nanoparticle Based Semi-Automated Antibody Phage Display Biopanning

Angela Chiew Wen Ch’ng, Azimah Ahmad, Zoltán Konthur, and Theam Soon Lim

19. A Binding Potency Assay for Pritumumab and Ecto-Domain Vimentin 

Ivan Babic, Santosh Kesari, and Mark C. Glossy

20. A Method to Detect the Binding of Hyper-Glycosylated Fragment Crystallizable (Fc) Region of Human IgG1 to Glycan Receptors

Patricia Blundell and Richard Pleass

21. A Cell-Based Reporter Assay Measuring the Activation of Fc Gamma Receptors Induced by Therapeutic Monoclonal Antibodies

Michihiko Aoyama, Minoru Tada, and Akiko Ishii-Watabe

22. “BIClonals”: Production of Bispecific Antibodies in IgG Format in Transiently Transfected Mammalian Cells

Dana Litvak-Greenfeld, Lilach Vaks, Stav Dror, Limor Nahary, and Itai Benhar

23. Production of Stabilized Antibody Fragments in the E. coli Bacterial Cytoplasm and in Transiently-Transfected Mammalian Cells

Racheli Birnboim-Perach, Yehudit Grinberg, Lilach Vaks, Limor Nahary, and Itai Benhar