Antibody engineering : methods and protocols


    • Lo, Benny K. C.


Antibody engineering : methods and protocols

edited by Benny K.C. Lo

(Methods in molecular biology / John M. Walker, series editor, v. 248)

Humana Press, c2004

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Includes bibliographical references and index



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.


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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