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  • Phage display of peptides in ligand selection for use in affinity chromatography.

Phage display of peptides in ligand selection for use in affinity chromatography.

Methods in molecular biology (Clifton, N.J.) (2008-10-02)
Joanne L Casey, Andrew M Coley, Michael Foley
ABSTRACT

Large repertoires of peptides displayed on bacteriophage have been extensively used to select for ligand-binding molecules. This is a relatively straightforward process involving several cycles of selection against target molecules, and the resulting ligands can be tailored to various applications. In this chapter we describe detailed methods to select peptide ligands for affinity chromatography, with particular focus on selection of peptides that mimic antigen epitopes. The selection process involves screening a phage peptide library against a monoclonal antibody, proving the peptide is an authentic epitope mimic and coupling the peptide mimotope to an affinity resin for purifying antibodies from human serum. There are several other applications of phage peptides that could be used for affinity chromatography; the approaches are outlined, but detailed methods have not been included.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Goat Anti-Human IgG Antibody, Fc, HRP conjugate, Chemicon®, from goat
Sigma-Aldrich
o-Phenylenediamine dihydrochloride, tablet, 5 mg substrate per tablet