Noncompetitive Homogeneous Detection of Small Molecules Using Synthetic Nanopeptamer-Based Luminescent Oxygen Channeling

Our group has previously developed immunoassays for noncompetitive detection of small molecules based on the use of phage borne anti-immunocomplex peptides. Recently, we substituted the phage particles by biotinylated synthetic anti-immunocomplex peptides complexed with streptavidin and named these...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2018-05, Vol.90 (10), p.6187-6192
Main Authors: Lassabe, Gabriel, Kramer, Karl, Hammock, Bruce D, González-Sapienza, Gualberto, González-Techera, Andrés
Format: Article
Language:English
Subjects:
Antibodies
Aptamers, Peptide - chemistry
Assaying
Atrazine
Atrazine - analysis
Automation
Beads
Channeling
Chemistry
Drinking water
Environmental monitoring
Herbicides
Immunoassays
Luminescence
Molecules
Nanoparticles
Occupational health
Oxygen
Oxygen - chemistry
Peptides
Pesticides
Phages
Rivers - chemistry
Signal generation
Streptavidin
Target detection
Triazines - analysis
Water analysis
Water Pollutants, Chemical - analysis
Water sampling
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Summary:Our group has previously developed immunoassays for noncompetitive detection of small molecules based on the use of phage borne anti-immunocomplex peptides. Recently, we substituted the phage particles by biotinylated synthetic anti-immunocomplex peptides complexed with streptavidin and named these constructs nanopeptamers. In this work, we report the results of combining AlphaLisa, a commercial luminescent oxygen channeling bead system, with nanopeptamers for the development of a noncompetitive homogeneous assay for the detection of small molecules. The signal generation of AlphaLisa assays relies on acceptor–donor bead proximity induced by the presence of the analyte (a macromolecule) simultaneously bound by antibodies immobilized on the surface of these beads. In the developed assay, termed as nanoAlphaLisa, bead proximity is sustained by the presence of a small model molecule (atrazine, MW = 215) using an antiatrazine antibody captured on the acceptor bead and an atrazine nanopeptamer on the donor bead. Atrazine is one of the most used pesticides worldwide, and its monitoring in water has relevant human health implications. NanoAlphaLisa allowed the homogeneous detection of atrazine down to 0.3 ng/mL in undiluted water samples in 1 h, which is 10-fold below the accepted limit in drinking water. NanoAlphaLisa has the intrinsic advantages for automation and high-throughput, simple, and fast homogeneous detection of target analytes that AlphaLisa assay provides.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.8b00657