UNIT 9.8 Phage Display Selection and Analysis of Ab-Binding Epitopes
Published Online: 1 NOV 2002
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Immunology
How to Cite
Enshell-Seijffers, D. and Gershoni, J. M. 2002. Phage Display Selection and Analysis of Ab-Binding Epitopes. Current Protocols in Immunology. 50:9.8:9.8.1–9.8.27.
- Published Online: 1 NOV 2002
- Published Print: AUG 2002
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The identification and characterization of B cell epitopes by combinatorial phage display peptide analyses is based on the principle that unique peptides can be affinity-purified from an enormous collection of random peptides. Moreover, once selected, the peptide sequence can be elucidated; filamentous bacteriophages have been genetically engineered to incorporate the DNA template corresponding to the peptide displayed on its surface. This unit begins with a discussion of some of the factors that distinguish available libraries. Protocols are then provided for affinity selection of antibody-specific phages, determination of phage titer, confirmation and amplification of positive phages, phage characterization, and construction of custom-tailored phages. The selection protocol in this unit is specific and designed for libraries that are used in the authors' laboratory and are based on the fth1 or fd-tet derived vectors. However, information is included for adapting these protocols to the specific requirements of other phage display libraries.