Part I: Introduction to Single-Domain Antibodies
1. Nanobodies: From Serendipitous Discovery of Heavy Chain-Only Antibodies in Camelids to a Wide Range of Useful Applications
Fangling Ji, Jun Ren, Cécile Vincke, Lingyun Jia, and Serge Muyldermans
2. Overview, Generation, and Significance of Variable New Antigen Receptors (VNARs) as a Platform for Drug and Diagnostic Development
Samata S. Pandey, Marina Kovaleva, Caroline J. Barelle, and Obinna C. Ubah
Part II: Isolation of Single-Domain Antibodies
3. Llama DNA Immunization and Isolation of Functional Single-Domain Antibody Binders Frédéric Trempe, Martin A. Rossotti, Tahir Maqbool, C. Roger MacKenzie, and Mehdi Arbabi-Ghahroudi
4. Preparation of Immune and Synthetic VNAR Libraries as Sources of High-Affinity Binders
Jahaziel Gasperin-Bulbarela, Olivia Cabanillas-Bernal, Salvador Dueñas, and Alexei F. Licea-Navarro
5. Isolation of Single-Domain Antibodies to Transmembrane Proteins Using Magnetized Yeast Cell Targets
Kaitlyn Bacon, Stefano Menegatti, and Balaji M. Rao
6. A Transgenic Heavy Chain IgG Mouse Platform as a Source of High Affinity Fully Human Single-Domain Antibodies for Therapeutic Applications Dubravka Drabek, Rick Janssens, Rien van Haperen, and Frank Grosveld
Part III: Expression of Single-Domain Antibodies
7. Cytoplasmic Production of Nanobodies and Nanobody-Based Reagents by Co-Expression of Sulfhydryl Oxidase and DsbC Isomerase
Ario de Marco
8. Small Scale Secretory VHH Expression in Saccharomyces cerevisiae Michiel M. Harmsen, Marga van Hagen-van Setten, and Peter T.J. Willemsen
9. Production of Single-Domain Antibodies in Pichia pastoris
Yusei Matsuzaki, Kaho Kajiwara, Wataru Aoki, and Mitsuyoshi Ueda
10. Production of Designer VHH-Based Antibodies in Plants
Henri De Greve
Part IV: Single-Domain Antibody Engineering and Characterization
11. Assessing the Aggregation Propensity of Single-Domain Antibodies Upon Heat-Denaturation Employing the ΔTm Shift
Patrick Kunz
12. Facile Affinity Maturation of Single-Domain Antibodies Using Next-Generation DNA Sequencing
Michael J. Lowden, Henk van Faassen, Shalini Raphael, Shannon Ryan, Greg Hussack, and Kevin A. Henry
13. Engineering pH-Sensitive Single-Domain Antibodies
Tosha M. Laughlin and James R. Horn
14. Humanization of Camelid Single-Domain Antibodies Traian Sulea
15. Creation of Multimeric Single-Domain Antibodies Using Bacterial Superglues
Paul J. Wichgers Schreur, Sandra van de Water, and Jeroen Kortekaas
16. Introducing Cysteines into Nanobodies for Site-Specific Labelling
Simon Boje Hansen and Kasper Røjkjær Andersen
17. Affinity-Guided Site Selective Labelling of Nanobodies With Aldehyde Handles
Anders Märcher and Kurt V. Gothelf
18. Cytoplasmic Expression of Nanobodies with Formylglycine Generating Enzyme Tag and Conversion to a Bio-Orthogonal Aldehyde Group
Da Li, Qiang Peng, Chungdong Huang, Berlin Zang, Jun Ren, Fangling Ji, Serge Muyldermans, and Lingyun Jia
19. Site-Specific Fluorescent Labelling, Single-Step Immunocytochemistry, and Delivery of Nanobodies into Living Cells
