Includes supplementary material: sn.pub/extras
The exquisite binding specificity of antibodies has made them valuable tools from the laboratory to the clinic. Since the deillegalscription of the murine hybridoma technology by Köhler 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®, http://imgt.cines.fr 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 Völkel, 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, Jürgen Grünberg, 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
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.
"...as 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
The exquisite binding specificity of antibodies has made them valuable tools from the laboratory to the clinic. Since the deillegalscription of the murine hybridoma technology by Köhler 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.
Antibody Sequence And Structure.- Internet Resources for the Antibody Engineer.- The Kabat Database and a Bioinformatics Example.- IMGT, The International ImMunoGeneTics Information System®, http://imgt.cines.fr.- Antibody Variable Regions.- Studying Antibody Conformations by Ultracentrifugation and Hydrodynamic Modeling.- Antibody-Lead Generation.- PCR Cloning of Human Immunoglobulin Genes.- Antibody Humanization by CDR Grafting.- Selection of Human Antibodies from Phage Display Libraries.- Production of Human Single-Chain Antibodies by Ribosome Display.- Production of Human Antibodies from Transgenic Mice.- Selection of Internalizing Antibodies for Drug Delivery.- Engineering Multivalent Antibody Fragments for In Vivo Targeting.- Production of Recombinant Bispecific Antibodies.- Antibody Expression And Optimization.- Expression and Isolation of Recombinant Antibody Fragments in E. coli.- Expression of Recombinant Antibodies in Mammalian Cell Lines.- Human Antibody Production Using Insect-Cell Expression Systems.- Antibody Production in Transgenic Plants.- Directed Mutagenesis of Antibody Variable Domains.- Antibody Affinity Maturation by Chain Shuffling.- Antibody Affinity Maturation by Random Mutagenesis.- Developing a Minimally Immunogenic Humanized Antibody by SDR Grafting.- Antibody Characterization and Novel Applications.- Antibody Purification by Column Chromatography.- Affinity Measurement Using Surface Plasmon Resonance.- Kinetic Exclusion Assays to Study High-Affinity Binding Interactions in Homogeneous Solutions.- Characterization of Antibody-Antigen Interactions by Fluorescence Spectroscopy.- Antibody Epitope Mapping Using Arrays of Synthetic Peptides.- Mapping Antibody:Antigen Interactions by Mass Spectrometry and Bioinformatics.- Radiometal Labeling ofAntibodies and Antibody Fragments for Imaging and Therapy.- Production and Characterization of Anti-Cocaine Catalytic Antibodies.- Recombinant Immunotoxins in the Treatment of Cancer.- Antibodies in Proteomics.- Targeting of Antibodies Using Aptamers.
"...as 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
Inhaltsverzeichnis
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®, http://imgt.cines.fr
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 Völkel, 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, Jürgen Grünberg, 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
Klappentext
A core collection of diverse 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 selection and generation of human recombinant antibodies by humanization approaches, molecular display technologies and transgenic animals. Procedures are also described on restructuring antibody leads into monovalent, multivalent, and bispecific binding fragments for a wide variety of in vivo applications. 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.
Includes supplementary material: sn.pub/extras
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