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Anal Chem. 2015 Dec 1;87(23):11660-5. doi: 10.1021/acs.analchem.5b00702. Epub 2015 Nov 11.

Aptamer-Phage Reporters for Ultrasensitive Lateral Flow Assays.

Author information

1
Department of Biology & Biochemistry, University of Houston , Houston, Texas 77004, United States.
2
Department of Chemical & Biomolecular Engineering, University of Houston , Houston, Texas 77204, United States.
3
Centro de Biotecnología FEMSA, Tecnológico de Monterrey , Monterrey, Nuevo León, Mexico.

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-biotin 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.

PMID:
26456715
PMCID:
PMC4666763
DOI:
10.1021/acs.analchem.5b00702
[Indexed for MEDLINE]
Free PMC Article

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