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by Dirk Saerens (Editor), Serge Muyldermans (Editor)
Part I: Overview of Single Domain Antibodies
1. From Whole Monoclonal Antibodies to Single Domain Antibodies: Think Small
Jean-Luc Teillaud
2. Introduction to Heavy Chain Antibodies and Derived Nanobodies
Cécile Vincke and Serge Muyldermans
3. Overview and Discovery of IgNARs and Generation of VNARs
Stewart D. Nuttall
Part II: Single Domain Antibody Library Construction
4. Creation of the Large and Highly Functional Synthetic Repertoire of Human VH and Vκ Domain Antibodies
Olga Ignatovich, Laurent Jespers, Ian M, Tomlinson, and Ruud M.T. de Wildt
5. Preparation of a Naïve Library of Camelid Single Domain Antibodies
Aurelien Olichon and Ario de Marco
Part III: Selection of Single Domain Antibodies
6. Selection by Phage Display of Single Domain Antibodies Specific to Antigens in Their Native Conformation
Peter Verheesen and Toon Laeremans
7. Semi-Automated Panning of Naive Camelidae Libraries and Selection of Single-Domain Antibodies Against Peptide Antigens
Jyothi Kumaran, Roger MacKenzie, and Mehdi Arbabi-Ghahroudi
8. Pichia Surface Display: A Tool for Screening Single Domain Antibodies
Kristof De Schutter and Nico Callewaert
9. Bacterial Two Hybrid: A Versatile One-Step Intracellular Selection Method
Mireille Pellis, Serge Muyldermans, and Cécile Vincke
10. Intracellular Antibody Capture (IAC) Methods for Single Domain Antibodies
Tomoyuki Tanaka and Terence H. Rabbitts
11. Selection of Functional Single Domain Antibody Fragments for Interfering with Protein-Protein Interactions Inside Cells: A "One Plasmid" Mammalian Two-Hybrid System
Tomoyuki Tanaka and Terence H. Rabbitts
12. Cell-Free Selection of Domain Antibodies by In Vitro Compartmentalization
Armin Sepp and Andrew Griffiths
13. Selection of VHHs Under Application Conditions
Edward Dolk, Theo Verrips, and Hans de Haard
14. Isolation and Characterization of Clostridium difficile Toxin-Specific Single-Domain Antibodies
Greg Hussack, Mehdi Arbabi-Ghahroudi, C. Roger MacKenzie, and Jamshid Tanha
15. Selection of VHH Antibody Fragments that Recognize Different Aβ Depositions Using Complex Immune Libraries
Rinse Klooster, Kim S. Rutger, and Sylvère van der Maarel
Part IV: Expression of Single Domain Antibodies and Derivatives
16. Expression of Single-Domain Antibodies in Bacterial Systems
Toya Nath Baral and Mehdi Arbabi-Ghahroudi
17. Expression of VHHs in S. cerevisiae
Andrea Gorlani, Hans de Haard, and Theo Verrips
18. Stable Expression of Chimeric Heavy Chain Antibodies in CHO Cells
Vishal Agrawal, Igor Slivac, Sylvie Perret, Louis Bisson, Gilles St-Laurent, Yanal Murad, Jianbing Zhang, and Yves Durocher
19. Production of Camel-Like Antibodies in Plants
Sylvie De Buck, Vikram Virdi, Thomas De Meyer, Kirsten De Wilde, Robin Piron, Jonah Nolf, Els Van Lerberge, Annelies De Paepe, and Ann Depicker
Part V: Improvement and Applications of Single Domain Antibodies
20. Selecting and Purifying Autonomous Human Variable Heavy (VH) Domains
Raffi Tonikian and Sachdev S. Sidhu
21. Solubility and Stability Engineering of Human VH Domains
Dae Young Kim, Wen Ding, and Jamshid Tanha
22. Improvement of Proteolytic Stability through In Silico Engineering
Lucy Rutten, Hans de Haard, and Theo Verrips
23. Selection of Human VH Single Domains with Improved Biophysical Properties by Phage Display
Kip Dudgeon, Romain Rouet, Kristoffer Famm, and Daniel Christ
Back Jacket
The development of the hybridoma technology created the possibility to obtain unlimited amounts of monoclonal antibodies (mAb) with high specificity and affinity for any target and to introduce mAbs in a wide range of applications; however, the bulky size of mAbs, costly production, and cumbersome engineering hampered regularly their streamlined development in some applications. In Single Domain Antibodies: Methods and Protocols, expert researchers examine single variable domain antibody fragments, referred to as VH, VL, VHH or VNAR. These fragments are the smallest intact antigen-binding fragments that can be produced recombinantly at low cost. Written in the highly successful Methods in Molecular Biology(TM) 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.
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