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Single Domain Antibodies By Dirk Saerens

Single Domain Antibodies by Dirk Saerens

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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;

Single Domain Antibodies Summary

Single Domain Antibodies: Methods and Protocols by Dirk Saerens

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.

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

Cecile 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 V Domain Antibodies

Olga Ignatovich, Laurent Jespers, Ian M, Tomlinson, and Ruud M.T. de Wildt

5. Preparation of a Naive 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 Cecile 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 A Depositions Using Complex Immune Libraries

Rinse Klooster, Kim S. Rutger, and Sylvere 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

Stewart D. Nuttall

Additional information

Single Domain Antibodies: Methods and Protocols by Dirk Saerens
Humana Press Inc.
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