title: Tagged polymerase chain reaction subtractive hybridization for the enrichment of phage display random peptide libraries.
abstract: Affinity selection of phage display peptide libraries is routinely used for isolating peptides capable of binding a range of molecules, including antibodies and receptors. This process is most successful when the selecting molecule is relatively pure, for example, a monoclonal antibody. However, isolation of peptides able to bind to target molecules present in a complex mixture is more difficult because the affinity selection process isolates peptides capable of binding to all molecules present in the mixture. Here we describe the development of a tagged polymerase chain reaction (PCR) subtractive hybridization method that is universally applicable for the targeted isolation of peptides able to bind to unique molecules within a complex mixture. We also describe a discriminatory limiting dilution PCR method that can be used to optimize hybridization conditions.
authors.affiliation: Institute of Infections, Immunity, and Inflammation, School of Molecular Medical Sciences, Queen's Medical Centre, University of Nottingham, Nottingham, NG7 2UH, UK.
authors.forename: Alexander W
authors.initials: AW
authors.lastname: Tarr
authors.forename: Steven P
authors.initials: SP
authors.lastname: Boneham
authors.forename: Anna M
authors.initials: AM
authors.lastname: Grabowska
authors.forename: Jonathan K
authors.initials: JK
authors.lastname: Ball
mesh_terms: D001435:Bacteriophages / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D006801:Humans; D006824:Hybridization, Genetic; D010446:Peptide Fragments / Q000235:genetics* / Q000378:metabolism; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D013382:Subtraction Technique
chemical_list: D010446:Peptide Fragments; D019151:Peptide Library