Aptamer-Phage Reporters for Ultrasensitive Lateral Flow Assays

Abstract

We introduce the modification of bacteriophage particles with aptamers for use as bioanalytical reporters, and demonstrate the use of these particles in ultrasensitive lateral flow assays. M13 phage displaying an in vivo biotinylatable peptide (AviTag) genetically fused to the phage tail protein pIII were used as reporter particle scaffolds, with biotinylated aptamers attached via avidin朾iotin linkages, and horseradish peroxidase (HRP) reporter enzymes covalently attached to the pVIII coat protein. These modified viral nanoparticles were used in immunochromatographic sandwich assays for the direct detection of IgE and of the penicillin-binding protein from Staphylococcus aureus (PBP2a). We also developed an additional lateral flow assay for IgE, in which the analyte is sandwiched between immobilized anti-IgE antibodies and aptamer-bearing reporter phage modified with HRP. The limit of detection of this LFA was 0.13 ng/mL IgE, ?100 times lower than those of previously reported IgE assays.

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Citation

Copyright 2015 Analytical Chemistry. This is a post-print version of a published paper that is available at: https://pubs.acs.org/doi/abs/10.1021/acs.analchem.5b00702. Recommended citation: Adhikari, Meena, Ulrich Strych, Jinsu Kim, Heather Goux, Sagar Dhamane, Mohan-Vivekanandan Poongavanam, Anna EV Hagstro?m, Katerina Kourentzi, Jacinta C. Conrad, and Richard C. Willson. "Aptamer-phage reporters for ultrasensitive lateral flow assays." Analytical chemistry 87, no. 23 (2015): 11660-11665. doi: 10.1021/acs.analchem.5b00702. This item has been deposited in accordance with publisher copyright and licensing terms with the author's permission.