Single Domain Antibodies. Methods and Protocols (Methods in Molecular Biology, Vol. 911)

Description

• Features methods of selection, expression, improvement, and library construction of single domain antibodies
• Includes step-by-step instruction that has been tested in labs around the world
• Provides key tips and implementation advice that is essential for success

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™ 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.

Content Level » Professional/practitioner
Keywords » Heavy chain antibodies - Single variable domain - V-NAR - VHH - sdAbs
Related subjects » Immunology

TABLE OF CONTENTS

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
• 24. Improvement of Single Domain Antibody Stability by Disulfide Bond Introduction
  Yoshihisa Hagihara and Dirk Saerens
• 25. Characterization of Single-Domain Antibodies with an Engineered Disulfide Bond
  Greg Hussack, C. Roger MacKenzie, and Jamshid Tanha
• 26. Affinity Maturation of Single-Domain Antibodies by Yeast Surface Display
  Akiko Koide and Shohei Koide
• 27. Multivalent Display of Single-Domain Antibodies
  Jianbing Zhang and C. Roger MacKenzie
• 28. Methods for Determining the PK Parameters of AlbudAbs and of Long Serum Half Life Drugs Made Using the AlbudAb Technology
  Daniel Rycroft and Lucy J. Holt
• 29. Fluorescent Protein Specific Nanotraps to Study Protein-Protein Interactions and Histone-Tail Peptide Binding
  Garwin Pichler, Heinrich Leonhardt, and Ulrich Rothbauer
• 30. Site-Specific Labeling of His-Tagged Nanobodies with 99mTc: A Practical Guide
  Catarina Xavier, Nick Devoogdt, Sophie Hernot, Ilse Vaneycken, Matthias D'Huyvetter, Jens De Vos, Sam Massa, Tony Lahoutte, and Vicky Caveliers
• 31. Nanobody-Based Chromatin Immunoprecipitation
  Trong Nguyen Duc, Gholamreza Hassanzadeh-Ghassabeh, Dirk Saerens, Eveline Peeters, Daniel Charlier, and Serge Muyldermans
• 32. User-Friendly Expression Plasmids Enable the Fusion of VHHs to Application-Specific Tags
  Ario de Marco
• 33. Application of Single Domain Antibodies in Tumor Histochemistry
  Kien T. Mai and C. Roger MacKenzie

Part VI: Case Studies

• 34. Nanobodies as Structural Probes of Protein Misfolding and Fibril Formation
  Erwin De Genst and Christopher M. Dobson
• 35. Molecular Imaging Using Nanobodies: A Case Study
  Nick Devoogdt, Catarina Xavier, Sophie Hernot, Ilse Vaneycken, Matthias D'Huyvetter, Jens De Vos, Sam Massa, Patrick De Baetselier, Vicky Caveliers, and Tony Lahoutte
• 36. Case Study on Live Cell Apoptosis-Assay Using Lamin-Chromobody Cell-Lines for High-Content Analysis
  Kourosh Zolghadr, Jacqueline Gregor, Heinrich Leonhardt, and Ulrich Rothbauer