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Single Domain Antibodies

Methods and Protocols

Dirk Saerens, Serge Muyldermans (Herausgeber)

Buch | Hardcover

580 Seiten

2012
Humana Press Inc. (Verlag)
978-1-61779-967-9 (ISBN)

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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; 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.

From Whole Monoclonal Antibodies to Single Domain Antibodies: Think Small.- Introduction to Heavy Chain Antibodies and Derived Nanobodies.- Overview and Discovery of IgNARs and Generation of VNARs.- Creation of the Large and Highly Functional Synthetic Repertoire of Human VH and Vκ Domain Antibodies.- Preparation of a Naïve Library of Camelid Single Domain Antibodies.- Selection by Phage Display of Single Domain Antibodies Specific to Antigens in Their Native Conformation.- Semi-Automated Panning of Naive Camelidae Libraries and Selection of Single-Domain Antibodies Against Peptide Antigens.- Pichia Surface Display: A Tool for Screening Single Domain Antibodies.- Bacterial Two Hybrid: A Versatile One-Step Intracellular Selection Method.- Intracellular Antibody Capture (IAC) Methods for Single Domain Antibodies.- Selection of Functional Single Domain Antibody Fragments for Interfering with Protein-Protein Interactions Inside Cells: A “One Plasmid” Mammalian Two-Hybrid System.- Cell-Free Selection of Domain Antibodies by In Vitro Compartmentalization.- Selection of VHHs Under Application Conditions.- Isolation and Characterization of Clostridium difficile Toxin-Specific Single-Domain Antibodies.- Selection of VHH Antibody Fragments that Recognize Different Aβ Depositions Using Complex Immune Libraries.- Expression of Single-Domain Antibodies in Bacterial Systems.- Expression of VHHs in S. cerevisiae.- Stable Expression of Chimeric Heavy Chain Antibodies in CHO Cells.- Production of Camel-Like Antibodies in Plants.- Selecting and Purifying Autonomous Human Variable Heavy (VH) Domains.- Solubility and Stability Engineering of Human VH Domains.- Improvement of Proteolytic Stability through In Silico Engineering.- Selection of Human VH Single Domains with Improved Biophysical Properties by Phage Display.- Improvement of Single Domain Antibody Stability by Disulfide Bond Introduction.- Characterization of Single-Domain Antibodies withan Engineered Disulfide Bond.- Affinity Maturation of Single-Domain Antibodies by Yeast Surface Display.- Multivalent Display of Single-Domain Antibodies.- Methods for Determining the PK Parameters of AlbudAbs and of Long Serum Half Life Drugs Made Using the AlbudAb Technology.- Fluorescent Protein Specific Nanotraps to Study Protein-Protein Interactions and Histone-Tail Peptide Binding.- Site-Specific Labeling of His-Tagged Nanobodies with 99mTc: A Practical Guide.- Nanobody-Based Chromatin Immunoprecipitation.- User-Friendly Expression Plasmids Enable the Fusion of VHHs to Application-Specific Tags.- Application of Single Domain Antibodies in Tumor Histochemistry.- Nanobodies as Structural Probes of Protein Misfolding and Fibril Formation.- Molecular Imaging Using Nanobodies: A Case Study.- Case Study on Live Cell Apoptosis-Assay Using Lamin-Chromobody Cell-Lines for High-Content Analysis.

Erscheint lt. Verlag	11.8.2012
Reihe/Serie	Methods in Molecular Biology ; 911
Zusatzinfo	35 Illustrations, color; 54 Illustrations, black and white; XV, 580 p. 89 illus., 35 illus. in color.
Verlagsort	Totowa, NJ
Sprache	englisch
Maße	178 x 254 mm
Themenwelt	Studium ► Querschnittsbereiche ► Infektiologie / Immunologie
ISBN-10	1-61779-967-X / 161779967X
ISBN-13	978-1-61779-967-9 / 9781617799679
Zustand	Neuware