Phage Display: A Laboratory Manual
Phage-display technology has begun to make critical contributions to the study of molecular recognition. DNA sequences are cloned into phage, which then present on their surface the proteins encoded by the DNA. Individual phage are rescued through interaction of the displayed protein with a ligand, and the specific phage is amplified by infection of bacteria. Phage-display technology is powerful but challenging and the aim of this manual is to provide comprehensive instruction in its...
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Phage-display technology has begun to make critical contributions to the study of molecular recognition. DNA sequences are cloned into phage, which then present on their surface the proteins encoded by the DNA. Individual phage are rescued through interaction of the displayed protein with a ligand, and the specific phage is amplified by infection of bacteria. Phage-display technology is powerful but challenging and the aim of this manual is to provide comprehensive instruction in its theoretical and applied so that any scientist with even modest molecular biology experience can effectively employ it. The manual reflects nearly a decade of experience with students of greatly varying technical expertise andexperience who attended a course on the technology at Cold Spring Harbor Laboratory. Phage-display technology is growing in importance and power. This manual is an unrivalled source of expertise in its execution and application. Trends in Genetics The book's raison d'etre is as a source of detailed protocols. These are clearly written, and include ideas for experimental controls and useful trouble-shooting notes. Jargon is generally avoided, and the protocols are up to date. [This] is a much-needed and well-executed attempt at theoretical and practical support to any researcher, established or novice, using phage display.
Sect. 1Phage Display1Filamentous Phage Biology2Phage-display Vectors3Antibody Libraries4Peptide Libraries5Functional Domains and Scaffolds6Gene Fragment Libraries and Genomic and cDNA Expression CloningSect. 2Antibody Libraries7Overview: Amplification of Antibody Genes8Generation of Antibody Libraries: Immunization, RNA Preparation, and cDNA Synthesis9Generation of Antibody Libraries: PCR Amplification and Assembly of Light- and Heavy-chain Coding Sequences10Selection from Antibody Libraries11Analysis of Selected Antibodies12Production and Purification of Fab and ScFv13Antibody EngineeringSect. 3Peptide Libraries14Overview: Peptide Libraries15General Phage Methods16Production of Peptide Libraries17Screening Peptide Libraries18Analysis of Phage-borne Peptides19Construction and Use of pIII-displayed Peptide LibrariesSect. 4Other Libraries and Other Methods of Panning20Construction and Selection from Gene Fragment Phage-display Expression Libraries21Construction and Selection from cDNA Phage-display Expression Libraries22In Vivo Selection of Phage-display Libraries23Cell-surface Selection and Analysis of Monoclonal Antibodies from Phage LibrariesApp. 1: Useful InformationApp. 2: RecipesApp. 3: General ProceduresApp. 4: CautionsApp. 5: SuppliersApp. 6: Trademarks
\ Trends in GeneticsThe book's raison d'etre is as a source of detailed protocols. These are clearly written, and include ideas for experimental controls and useful trouble-shooting notes. Jargon is generally avoided, and the protocols are up to date.…[This] is a much-needed and well-executed attempt at theoretical and practical support to any researcher, established or novice, using phage display.\ \ \ \ \ BooknewsPhage display is a rapidly growing field, but the technology has a reputation for requiring considerable technical expertise. This manual aims to provide comprehensive instruction in the theory and applications of phage-display technologies so that any scientist with even modest experience in molecular biology can employ them. In addition to an overview of key aspects of phage display, sections cover the construction, screening, and analysis of antibody libraries on the surface of phage; phage-displayed peptide libraries; and gene- fragment and cDNA-expression libraries. Appendices summarize commonly used experimental procedures, data, recipes, suppliers, and important precautions. Annotation c. Book News, Inc., Portland, OR (booknews.com)\ \
