TY - JOUR
T1 - Phage display selection and analysis of Ab-binding epitopes.
AU - Enshell-Seijffers, David
AU - Gershoni, Jonathan M.
PY - 2002/11
Y1 - 2002/11
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=43549089490&partnerID=8YFLogxK
U2 - 10.1002/0471142735.im0908s50
DO - 10.1002/0471142735.im0908s50
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C2 - 18432894
AN - SCOPUS:43549089490
SN - 1934-3671
VL - Chapter 9
SP - Unit 9.8
JO - Current Protocols in Immunology
JF - Current Protocols in Immunology
ER -