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

Tarr, Alexander W.; Boneham, Steven P.; Grabowska, Anna M.; Ball, Jonathan K.

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. © 2004 Elsevier Inc. All rights reserved.

Citation

Tarr, A. W., Boneham, S. P., Grabowska, A. M., & Ball, J. K. (2005). Tagged polymerase chain reaction subtractive hybridization for the enrichment of phage display random peptide libraries. Analytical Biochemistry, 339(1), 61-68. https://doi.org/10.1016/j.ab.2004.12.020

• Journal Article Type: Article
• Online Publication Date: Jan 12, 2005
• Publication Date: Apr 1, 2005
• Deposit Date: Nov 24, 2022
• Journal: Analytical Biochemistry
• Print ISSN: 0003-2697
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 339
• Issue: 1
• Pages: 61-68
• DOI: https://doi.org/10.1016/j.ab.2004.12.020
• Public URL: https://nottingham-repository.worktribe.com/output/3129686
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S0003269704010036?via%3Dihub