A1 Journal article – refereed
Luminometric Nanoparticle-Based Assay for High Sensitivity Detection of β-Amyloid Aggregation

List of Authors: Sari Pihlasalo, Takahiro Deguchi, Maria Virtamo, Jenna Jacobino, Karthik Chary, Francisco R. López-Picón, Gerda Brunhofer-Bolzer, Roope Huttunen, Adyary Fallarero, Pia Vuorela, Harri Härmä
Publication year: 2017
Journal: Analytical Chemistry
Journal name in source: Analytical chemistry
Journal acronym: Anal Chem
Volume number: 89
Issue number: 4
ISSN: 1520-6882
eISSN: 1520-6882

A nanoparticle-based assay utilizing time-resolved luminescence resonance energy transfer (TR-LRET) was developed for the detection of β-amyloid aggregation. The assay is based on the competitive adsorption of the sample and the acceptor-labeled protein to donor europium(III) polystyrene nanoparticles. The performance of the assay was demonstrated by following the fibrillization of β-amyloid peptide 1-42 (Aβ42) as a function of time and by comparing to the reference methods atomic force microscopy (AFM) and thioflavin T (ThT) assay. The fibrillization leads to reduced adsorption of Aβ42 to the nanoparticles increasing the TR-LRET signal. The investigated methods detected fibril formation with equal sensitivities. Eight potential fibrillization inhibitor compounds reported in the literature were tested and the results obtained with each method were compared. It was shown with AFM imaging that the inhibition of fibril formation was not complete with any of the compounds. The developed TR-LRET nanoparticle assay gave corresponding results with the AFM imaging. However, the ThT assay led to contradictory results, as low fluorescence signal was measured in the presence of all tested compounds suggesting inhibition of fibrillization. Our results suggest that the developed TR-LRET nanoparticle assay can be exploited for screening of potential β-amyloid aggregation inhibitors, whereas some of the tested compounds may be measured as false positive inhibitors with the much-utilized ThT assay.

Last updated on 2019-21-08 at 20:41