Antibody Engineering

Antibody Engineering : Methods and Protocols

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The exquisite binding specificity of antibodies has made them valuable tools from the laboratory to the clinic. Since the description of the murine hybridoma technology by Koehler and Milstein in 1975, a phenomenal number of mo- clonal antibodies have been generated against a diverse array of targets. Some of these have become indispensable reagents in biomedical research, while others were developed for novel therapeutic applications. The attractiveness of an- bodies in this regard is obvious-high target specificity, adaptability to a wide range of disease states, and the potential ability to direct the host's immune s- tem for a therapeutic response. The initial excitement in finding Paul Ehrlich's "magic bullet," however, was met with widespread disappointment when it was demonstrated that murine antibodies frequently elicit the human anti-murine an- body (HAMA) response, thus rendering them ineffective and potentially unsafe in humans. Despite this setback, advances in recombinant DNA techniques over the last 15-20 years have empowered the engineering of recombinant antibodies with desired characteristics, including properties to avoid HAMA. The ability to p- duce bulk quantities of recombinant proteins from bacterial fermentation also fueled the design of numerous creative antibody constructs. To date, the United States Food and Drug Administration has approved more than 10 recombinant antibodies for human use, and hundreds more are in the development pipeline. The recent explosion in genomic and proteomic information appears ready to deliver many more disease targets amenable to antibody-based therapy.
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Product details

  • Hardback | 562 pages
  • 160 x 230 x 30mm | 961.62g
  • Totowa, NJ, United States
  • English
  • 2004 ed.
  • XIV, 562 p.
  • 1588290921
  • 9781588290922
  • 2,617,378

Back cover copy

With 10 recent FDA approvals of therapeutic antibody drugs and hundreds more in the biotechnology pipeline, it is clear that engineered antibodies will play a major role in treating human diseases in the 21st century. In Antibody Engineering: Methods and Protocols, leading researchers and clinicians present a core collection of cutting-edge techniques for the generation, expression, optimization, and characterization of recombinant antibodies. Readily reproducible protocols for lead generation range from the cloning of human immunoglobulin genes to the generation of human recombinant antibodies by humanization approaches, molecular display technologies, and transgenic animals. The techniques for antibody expression and optimization utilize not only bacterial and mammalian cell cultures, but also insect cell cultures and transgenic plants. State-of-the-art technologies are described for the characterization of antigen-binding affinity and specificity with novel applications in radioimmunotargeting, cancer immunotherapy, drug abuse, and proteomics. Each fully tested protocol is described in step-by-step detail and includes background material, equipment and reagent lists, and tips on troubleshooting and avoiding pitfalls.
Comprehensive and up-to-date, Antibody Engineering: Methods and Protocols provides both laboratory and clinical investigators with a diverse array of the fundamental techniques upon which new and imaginative drug technologies can be successfully developed.
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Table of contents

Part I. Antibody Sequence and Structure

Internet Resources for the Antibody Engineer
Benny K. C. Lo and Yu Wai Chen

The Kabat Database and a Bioinformatics Example
George Johnson and Tai Te Wu

IMGT, The International ImMunoGeneTics Information System (R),
Marie-Paule Lefranc

Antibody Variable Regions: Toward a Unified Modeling Method
Nicholas Whitelegg and Anthony R. Rees

Studying Antibody Conformations by Ultracentrifugation and Hydrodynamic Modeling
Stephen E. Harding, Emma Longman, Beatriz Carrasco, Alvaro Ortega, and Jose Garcia de la Torre

Part II. Antibody-Lead Generation

PCR Cloning of Human Immunoglobulin Genes
James D. Marks and Andrew Bradbury

Antibody Humanization by CDR Grafting
Benny K. C. Lo

Selection of Human Antibodies from Phage Display Libraries
James D. Marks and Andrew Bradbury

Production of Human Single-Chain Antibodies by Ribosome Display
Mingyue He, Neil Cooley, Alison Jackson, and Michael J. Taussig

Production of Human Antibodies from Transgenic Mice
C. Geoffrey Davis, Xiao-Chi Jia, Xiao Feng, and Mary Haak-Frendscho

Selection of Internalizing Antibodies for Drug Delivery
James D. Marks

Engineering Multivalent Antibody Fragments for In Vivo Targeting
Anna M. Wu

Production of Recombinant Bispecific Antibodies
Roland E. Kontermann, Tina Voelkel, and Tina Korn

Part III. Antibody Expression and Optimization

Expression and Isolation of Recombinant Antibody Fragments in E. coli
Keith A. Charlton

Expression of Recombinant Antibodies in Mammalian Cell Lines
Paul J. Yazaki and Anna M. Wu

Human Antibody Production Using Insect-Cell Expression Systems
Mary C. Guttieri and Mifang Liang

Antibody Production in Transgenic Plants
Eva Stoger, StefanSchillberg, Richard M. Twyman, Rainer Fischer, and Paul Christou

Directed Mutagenesis of Antibody Variable Domains
Kevin Brady and Benny K. C. Lo

Antibody Affinity Maturation by Chain Shuffling
James D. Marks

Antibody Affinity Maturation by Random Mutagenesis
Ikuo Fujii

Developing a Minimally Immunogenic Humanized Antibody by SDR Grafting
Syed V. S. Kashmiri, Roberto De Pascalis, and Noreen R. Gonzales

Part IV. Antibody Characterization and Novel Applications

Antibody Purification by Column Chromatography
Andrea Murray, C. Rosamund L. Graves, Kevin Brady, and Benny K. C. Lo

Affinity Measurement Using Surface Plasmon Resonance
Robert Karlsson and Anita Larsson

Kinetic Exclusion Assays to Study High Affinity Binding Interactions in Homogeneous Solutions
Robert C. Blake II and Diane A. Blake

Characterization of Antibody-Antigen Interactions by Fluorescence Spectroscopy
Sotiris Missailidis and Kevin Brady

Antibody Epitope Mapping Using Arrays of Synthetic Peptides
Ulrich Reineke

Mapping Antibody:Antigen Interactions by Mass Spectrometry and Bioinformatics
Alexandra Huhalov, Daniel I. R. Spencer, and Kerry A. Chester

Radiometal Labeling of Antibodies and Antibody Fragments for Imaging and Therapy
Ilse Novak-Hofer, Robert Waibel, Kurt Zimmermann, Roger Schibli, Jurgen Grunberg, Kerry A. Chester, Andrea Murray, Benny K. C. Lo, Alan C. Perkins, and P. August Schubiger

Production and Characterization of Anti-Cocaine Catalytic Antibodies
Paloma de Prada and Donald W. Landry

Recombinant Immunotoxins in the Treatment of Cancer
Ira Pastan, Richard Beers, and Tapan K. Bera

Antibodies in Proteomics
Andrew R. M. Bradbury, Nileena Velappan, Vittorio Verzillo, Milan Ovecka, Roberto Marzari, Daniele Sblattero, Leslie Chasteen, Robert Siegel, and Peter
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Review Text

" the book provides as comprehensive collection of clear, detailed methods it is very likely to prove its worth as a laboratory reference." - Immunology News

"...a valuable addition to in-lab books" - Microbiology Today
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Review quote

" the book provides as comprehensive collection of clear, detailed methods it is very likely to prove its worth as a laboratory reference." - Immunology News

"...a valuable addition to in-lab books" - Microbiology Today
show more