Non-ionic detergents facilitate non-specific binding of M13 bacteriophage to polystyrene surfaces

Hakami, Abdulrahim R.; Ball, Jonathan K.; Tarr, Alexander W.

Abstract

© 2015 Elsevier B.V. Phage-displayed random peptide libraries are widely used for identifying peptide interactions with proteins and other substrates. Selection of peptide ligands involves iterative rounds of affinity enrichment. The binding properties of the selected phage clones are routinely tested using immunoassay after propagation to high titre in a bacterial host and precipitation using polyethylene glycol (PEG) and high salt concentration. These immunoassays can suffer from low sensitivity and high background signals. Polysorbate 20 (Tween® 20) is a non-ionic detergent commonly used in immunoassay washing buffers to reduce non-specific binding, and is also used as a blocking reagent. We have observed that Tween 20 enhances non-specific M13 library phage binding in a peptide-independent manner. Other non-ionic detergents were also found to promote significant, dose-dependent non-specific phage binding in ELISA. This effect was not observed for assays using phage concentrated by ultracentrifugation, suggesting that interactions occur between detergents and the PEG-precipitated phage, irrespective of the displayed peptide motif. This artefact may impact on successful affinity selection of peptides from phage-display libraries. We propose alternative methods for screening phage libraries for identifying binding interactions with target ligands.

Citation

Hakami, A. R., Ball, J. K., & Tarr, A. W. (2015). Non-ionic detergents facilitate non-specific binding of M13 bacteriophage to polystyrene surfaces. Journal of Virological Methods, 221, 1-8. https://doi.org/10.1016/j.jviromet.2015.04.023

• Journal Article Type: Article
• Acceptance Date: Apr 20, 2015
• Online Publication Date: Apr 27, 2015
• Publication Date: Sep 1, 2015
• Deposit Date: Jan 28, 2020
• Journal: Journal of Virological Methods
• Print ISSN: 0166-0934
• Electronic ISSN: 1879-0984
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 221
• Pages: 1-8
• DOI: https://doi.org/10.1016/j.jviromet.2015.04.023
• Public URL: https://nottingham-repository.worktribe.com/output/1860724
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S0166093415001676
• Additional Information: This article is maintained by: Elsevier; Article Title: Non-ionic detergents facilitate non-specific binding of M13 bacteriophage to polystyrene surfaces; Journal Title: Journal of Virological Methods; CrossRef DOI link to publisher maintained version: https://doi.org/10.1016/j.jviromet.2015.04.023; Content Type: article; Copyright: Copyright © 2015 Elsevier B.V. All rights reserved.