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Tagged polymerase chain reaction subtractive hybridization for the enrichment of phage display random peptide libraries

Lookup NU author(s): Dr Steve Boneham


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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. © 2004 Elsevier Inc. All rights reserved.

Publication metadata

Author(s): Tarr AW, Boneham SP, Grabowska AM, Ball JK

Publication type: Article

Publication status: Published

Journal: Analytical Biochemistry

Year: 2005

Volume: 339

Issue: 1

Pages: 61-68

Print publication date: 01/04/2005

Online publication date: 12/01/2005

ISSN (print): 0003-2697

ISSN (electronic): 1096-0309

Publisher: Elsevier


DOI: 10.1016/j.ab.2004.12.020

PubMed id: 15766711


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