Noncompetitive Homogeneous Detection of Small Molecules Using Synthetic Nanopeptamer-Based Luminescent Oxygen Channeling
| Autor: | Gabriel Lassabe, Bruce D. Hammock, Gualberto González-Sapienza, Karl Kramer, Andrés González-Techera |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Streptavidin Analyte Luminescence Aptamer Chemical Aptamers 01 natural sciences Article Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound Rivers Molecule Water Pollutants Triazines 010401 analytical chemistry Small molecule Combinatorial chemistry Acceptor 0104 chemical sciences Oxygen 030104 developmental biology chemistry Biotinylation Peptide Atrazine Other Chemical Sciences Aptamers Peptide Water Pollutants Chemical Macromolecule |
| Zdroj: | Analytical chemistry, vol 90, iss 10 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.8b00657 |
| Popis: | 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 immobilised 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 anti-atrazine 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 to 0.3 ng/mL in undiluted water samples in one hour, which is ten-fold below the accepted limit in drinking water. NanoAlphaLisa has the intrinsic advantages for automation, high-throughput, simple, and fast homogeneous detection of target analytes that AlphaLisa assay provides. |
| Databáze: | OpenAIRE |
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