Beschreibung
Monoclonal Antibodies: Methods and Protocols, Second Edition expands upon the previous edition with current, detailed modern approaches to isolate and characterize monoclonal antibodies against carefully selected epitopes. This edition includes new chapters covering the key steps to generate high quality monoclonals via different methods, from antigen generation to epitope mapping and quality control of the purified IgG. Chapters are divided into four parts corresponding to four distinct objectives. Part I covers monoclonal antibody generation, Part II deals with monoclonal antibody expression and purification, Part III presents methods for monoclonal antibody characterization and modification, and Part IV describes selected applications of monoclonal antibodies. 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.Authoritative and practical, Monoclonal Antibodies: Methods and Protocols, Second Edition provides crucial initial steps of monoclonal antibody generation and characterization with state-of-the art protocols.
Autorenporträt
InhaltsangabePart I: Antibody Generation1. Antigen Production for Monoclonal Antibody GenerationGiovanni Magistrelli, Pauline Malinge2. Method for the Generation of Antibodies Specific for Site- and Post-translational ModificationsHidemasa Goto and Masaki Inagaki3. Immunization, Hybridoma Generation and Selection for Monoclonal Antibody ProductionHyung-Yong Kim, Alexander Stojadinovic, and Mina J. Izadjoo4. Hybridoma Technology for the Generation of Rodent mAbs via Classical Fusion Efthalia Chronopoulou, Alejandro Uribe-Benninghoff, Cindi R. Corbett, and Jody D. Berry5. Generation of Rabbit Monoclonal AntibodiesPi-ChenYam and Katherine L. Knight6. Screening Hybridomas for Cell Surface Antigens by High-throughput Homogenous Assay and Flow CytometryAlejandro Uribe-Benninghoff, Teresa Cabral, Efthalia Chronopoulou, Jody D. Berry and Cindi R. Corbett7. Screening and Subcloning of High Producer Transfectomas using Semi-solid Media and Automated Colony PickerSuba Dharshanan and Cheah Swee Hung8. Design and Generation of Synthetic Antibody Libraries for Phage DisplayGang Chen and Sachdev S. Sidhu9. Selection and Screening using Antibody Phage Display LibrariesPatrick Koenig and Germaine Fuh10. Yeast Surface Display for Antibody Isolation: Library Construction, Library Screening, and Affinity MaturationJames A. Van Deventer and Karl Dane Wittrup11. Human B Cell Immortalization for Monoclonal Antibody ProductionJoyce Hui-Yuen, Siva Koganti,and Sumita Bhaduri-McIntosh12. Using Next-generation Sequencing for Discovery of High-frequency Monoclonal Antibodies in the Variable Gene Repertoires from Immunized MiceUlrike Haessler and Sai T. ReddyPart II: Antibody Expression and Purification13. Cloning, Reformatting, and Small Scale Expression of Monoclonal Antibody Isolated from Mouse, Rat, or Hamster HybridomaJeremy Loyau and François Rousseau 14. Cloning of Recombinant Monoclonal Antibodies from Hybridomas in a Single Mammalian Expression PlasmidNicole Müller-Sienerth, Cécile Crosnier, Gavin J. Wright and Nicole Staudt15. Monoclonal Antibody Purification by Ceramic Hydroxyapatite ChromatographyLarry J. Cummings, Russell G. Frost and Mark A. Snyder16. Rapid Purification of Monoclonal Antibodies using Magnetic MicrospheresPauline Malinge, Giovanni Magistrelli17. Generation of Cell Lines for Monoclonal Antibody ProductionKrista Alvin and Jianxin Ye18. Expression and Purification of Recombinant Antibody Formats and Antibody Fusion ProteinsMartin Siegemund, Fabian Richter, Oliver Seifert, Felix Unverdorben and Roland E. Kontermann19. Purification of Antibodies and Antibody Fragments using CaptureSelect Affinity ResinsPim Hermans, Hendrik Adams, Frank Detmers20. Reformatting of scFv antibodies into the scFv-Fc format and their Downstream PurificationEmil Bujak, Mattia Matasci, Dario Neri, and Sarah WulhfardPart III: Anitbody Characterization and Modification21. Antibody V and C Domain Sequence, Structure, and Interaction Analysis with Special Reference to IMGT®Eltaf Alamyar, Véronique Giudicelli, Patrice Duroux and Marie-Paule Lefranc22. Measuring Antibody Affinities as well as the Active Concentration of Antigens Present on a Cell SurfacePalaniswami Rathanaswami23. Determination of Antibody StructuresRobyn L. Stanfield24. Affinity Maturation of Monoclonal Antibodies by Multi Site-Directed MutagenesisHyung-Yong Kim, Alexander Stojadinovic, and Mina J. Izadjoo25. Epitope Mapping with Membrane-bound Synthetic Overlapping PeptidesTerumi Midoro-Horiuti and Randall M. Goldblum26. Epitope Mapping by Epitope Excision, Hydrogen/Deuterium Exchange and
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