Phage display : a laboratory manual
Author(s)
Bibliographic Information
Phage display : a laboratory manual
Cold Spring Harbor Laboratory Press, c2001
- : hbk
- : pbk
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Includes bibliographical references and index
Description and Table of Contents
Description
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 and experience who attended a course on the technology at Cold Spring Harbor Laboratory.
Table of Contents
- Phage display: filamentous phage biology
- phage-display vectors
- antibody libraries
- peptide libraries
- functional domains and scaffolds
- gene fragment libraries and genomic and cDNA expression cloning. Antibody libraries: overview - amplification of antibody genes
- generation of antibody libraries - immunization, RNA preparation and cDNA synthesis
- generation of antibody libraries - PCR amplification and assembly of light- and heavy-chain coding sequences
- selection from antibody libraries
- analysis of selected antibodies
- production and purification of fab and scFv
- antibody engineering. Peptide libraries: overview - peptide libraries
- general phage methods
- production of peptide libraries
- screening peptide libraries
- analysis of phage-borne peptides
- construction and use of pIII-displayed peptide libraries. Other libraries and other methods of panning: construction and selection from gene fragment phage-display expression libraries
- construction and selection from cDNA phage-display expression libraries
- in-vivo selection of phage-display libraries
- cell-surface selection and analysis of monoclonal antibodies from phage libraries. Appendices: useful information
- recipes
- general procedures
- cautions
- suppliers
- trademarks.
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