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Antibody Phage Display: Methods and Protocols: Methods in Molecular Biology, cartea 178

Editat de Philippa M. O’Brien, Robert Aitken
en Limba Engleză Paperback – 28 dec 2001
The closing years of the 19th century and the start of the 20th century witnessed the emergence of microbiology and immunology as discrete sci- tific disciplines, and in the work of Roux and Yersin, perhaps the first benefits of their synergy—immunotherapy against bacterial infection. As we advance into the new millennium, microbiology and immunology again offer a c- ceptual leap forward as antibody phage display gains increasing acceptance as the definitive technology for monoclonal production and unleashes new - portunities in immunotherapy, drug discovery, and functional genomics. In assembling Antibody Phage Display: Methods and Protocols, we have aimed to produce a resource of real value for scientists who have followed the development of phage display technology over the past decade. The founding principles of phage display have always held an elegant simplicity. We hope that readers will find similar clarity in the technical guidance offered by the book’s contributors. In meeting our objectives, we have tried to cover the broad scope of the technology and the key areas of library construction, scre- ing, antibody modification, and expression. Of course, the technology cont- ues to advance apace, but we trust that readers will be able to gage the potential of phage display from our coverage, that some of its subtleties will emerge, and that our selection of methods will prove appealing. We are indebted to all the contributing authors for sharing their expertise with the wider scientific community.
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Specificații

ISBN-13: 9780896037113
ISBN-10: 0896037118
Pagini: 401
Ilustrații: XV, 401 p.
Dimensiuni: 155 x 235 x 30 mm
Greutate: 0.77 kg
Ediția:2002
Editura: Humana Press Inc.
Colecția Humana
Seria Methods in Molecular Biology

Locul publicării:Totowa, NJ, United States

Public țintă

Research

Cuprins

Overview of Antibody Phage-Display Technology and Its Applications.- Standard Protocols for the Construction of Fab Libraries.- Standard Protocols for the Construction of scFv Libraries.- Broadening the Impact of Antibody Phage Display Technology.- Construction of Large Naïve Fab Libraries.- Construction of Polyclonal Antibody Libraries Using Phage Display.- Antigen-Driven Stimulation of B-Lymphocytes In Vitro.- 8 The Recovery of Immunoglobulin Sequences from Single Human B Cells by Clonal Expansion.- Panning of Antibody Phage-Display Libraries.- Selection of Antibodies Against Biotinylated Antigens.- Isolation of Anti-Hapten Specific Antibody Fragments from Combinatorial Libraries.- Blocking Immunodominant Epitopes by Competitive Deselection.- Rescue of a Broader Range of Antibody Specificities Using an Epitope-Masking Strategy.- Screening of Phage-Expressed Antibody Libraries by Capture Lift.- Antibody-Guided Selection Using Capture-Sandwich ELISA.- Proximity-Guided (ProxiMol) Antibody Selection.- Isolation of Human Monoclonal Antibodies Using Guided Selection with Mouse Monoclonal Antibodies.- Selecting Antibodies to Cell-Surface Antigens Using Magnetic Sorting Techniques.- Isolation of Human Tumor-Associated Cell Surface Antigen-Binding scFvs.- Subtractive Isolation of Single-Chain Antibodies Using Tissue Fragments.- Selection of Antibodies Based on Antibody Kinetic Binding Properties.- Selection of Functional Antibodies on the Basis of Valency.- Two-Step Strategy for Alteration of Immunoglobulin Specificity by In Vitro Mutagenesis.- Targeting Random Mutations to Hotspots in Antibody Variable Domains for Affinity Improvement.- Error-Prone Polymerase Chain Reaction for Modification of scFvs.- Use of Escherichia coli Mutator Cells to Mature Antibodies.- ChainShuffling to Modify Properties of Recombinant Immunoglobulins.- Generation of Bispecific and Tandem Diabodies.- High-Level Periplasmic Expression and Purification of scFvs.- Periplasmic Expression and Purification of Recombinant Fabs.- Expression of Antibody Fragments in Pichia pastoris.- Expression of VHH Antibody Fragments in Saccharomyces cerevisiae.- Intrabodies.- 34 Expression of scFvs and scFv Fusion Proteins in Eukaryotic Cells.- 35 Expression of Antibody Fab Fragments and Whole Immunoglobulin in Mammalian Cells.

Recenzii

"...this book lives up to its promises. It starts with an excellent introduction by Hennie Hoogenboom, reprinted from his Immunotechnology article, which sets the scene and provides an overview of the field for new readers...Members of the laboratory that used the book for reference found it very clear and all were keen that the review was completed as quickly as possible so that the book was returned to the laboratory where they could make use of it! I think this in itself supports the fact that the book is a useful volume." - Immunology News

"As with other volumes in this popular series, each protocol is discussed in detail, including an introduction, materials, methods notes, and references. It is a good purchase for research libraries where monoclonal antibodies are studied." E-Streams

Textul de pe ultima copertă

Antibody phage display, the definitive technology for monoclonal production, has now advanced to a stage where it can be performed in nonspecialized research laboratories. In Antibody Phage Display: Methods and Protocols, Philippa M. O'Brien and Robert Aitken combine in one volume a comprehensive collection of established antibody phage display protocols, each accompanied by authoritative guidance that will enable the nonspecialist to carry them out successfully. Coverage spans the construction of antibody libraries, the selection of antibody clones with the desired properties, and their modification, expression, and purification. Each readily reproducible method is described by a hands-on expert in step-by-step detail and includes a wealth of practical advice not found in the scientific literature. Extensive notes discuss pitfalls to avoid and offer many possible alternative methods to suit special research situations. An overview by one of the world's leading experts in antibody phage display, Dr. Hennie R. Hoogenboom, surveys the current status of the field and the future of the technology.
Comprehensive and highly practical, Antibody Phage Display: Methods and Protocols provides biochemists, molecular biologists, and immunologists with a gold-standard reference guide to the successful isolation, modification, and expression of recombinant antibodies using today's powerful phage display technology.

Caracteristici

Includes supplementary material: sn.pub/extras