Single-Domain Antibodies: Methods and Protocols

ISBN-13: 9781071620779 / Angielski / Miękka / 2022 / 612 str.

Greg Hussack; Kevin A. Henry

Opis

This volume covers current and emerging techniques for studying single-domain antibodies (sdAbs). Chapters guide readers through the biology and immunology of sdAbs in camelids and sharks, isolation of sdAbs, protein engineering approaches to optimize the solubility, stability, valency and antigen binding affinity of sdAbs, and specialized applications of sdAbs. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Single-Domain Antibodies: Methods and Protocols aims to be a useful, practical guide to help researchers further their studies in this field.

Spis treści

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 ΔT_m 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

Jan Gettemans

 

20.  Design and Validation of Site-Specifically Labelled Single-Domain Antibody-Based Tracers for In Vivo Fluorescence Imaging and Image-Guided Surgery

Noemi B. Declerck, Lukasz Mateusiak, and Sophie Hernot

 

21.  Design and Preparation of Photobodies: Light-Activated Single-Domain Antibody Fragments

Zahide Yilmaz, Benedikt Jedlitzke, and Henning D. Mootz

 

Part V: Applications of Single-Domain Antibodies

22.  Visualizing Filoviral Nucleoproteins Using Nanobodies Fused to the Ascorbate Peroxidase Derivatives APEX2 and dEAPX

Laura Jo Sherwood and Andrew Hayhurst

 

23.  Development of Glypican-2 Targeting Single-Domain Antibody CAR T Cells for Neuroblastoma

Nan Li and Mitchell Ho

 

24.  Single-Domain Antibodies for Intracellular Toxin Neutralization

Timothy F. Czajka and Nicholas J. Mantis

 

25.  Generation of Single-Domain Antibody-Based Recombinant Immunotoxins

Bryan D. Fleming and Mitchell Ho

 

26.  X-Ray Crystal Structure Analysis of VHH-Protein Antigen Complexes

Angham M. Ahmed and Cory L. Brooks

 

27.  Functionalization of Magnetic Beads with Biotinylated Nanobodies for MALDI-TOF/MS-Based Quantitation of Small Analytes

Gabriel Lassabe, Macarena Pírez-Schirmer, and Gualberto González-Sapienza

 

28.  Nanobody-Based Assays for the Detection of Environmental and Agricultural Contaminants

Feng Wang and Hong Wang

 

29.  Peptide Tag-Specific Nanobodies for Studying Proteins in Live Cells

Funmilayo O. Fagbadebo and Ulrich Rothbauer

 

30.  Nanobody-Based GFP Traps to Study Protein Localization and Function in Developmental Biology

Shinya Matsuda, Gustavo Aguilar, M. Alessandra Vigano, and Markus Affolter

 

31.  Optogenetic Activation of Intracellular Nanobodies

Daseuli Yu and Won Do Heo