Title |
A digital enzyme-linked immunosorbent assay for ultrasensitive measurement of amyloid-β 1–42 peptide in human plasma with utility for studies of Alzheimer’s disease therapeutics
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Published in |
Alzheimer's Research & Therapy, December 2016
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DOI | 10.1186/s13195-016-0225-7 |
Pubmed ID | |
Authors |
Linan Song, D. Richard Lachno, David Hanlon, Adam Shepro, Andreas Jeromin, Dipika Gemani, Jayne A. Talbot, Margaret M. Racke, Jeffrey L. Dage, Robert A. Dean |
Abstract |
Amyloid-β 1-42 peptide (Aβ1-42) is associated with plaque formation in the brain of patients with Alzheimer's disease (AD). Pharmacodynamic studies of AD therapeutics that lower the concentrations of Aβ1-42 in peripheral blood require highly sensitive assays for its measurement. A digital enzyme-linked immunosorbent assay (ELISA) using single molecule array (Simoa) technology has been developed that provides improved sensitivity compared with conventional ELISA methods using the same antibody reagents. A sensitive digital ELISA for measurement of Aβ1-42 using antibodies 3D6 and 21F12 was developed. Assay performance was evaluated by repeated testing of pooled human plasma and buffer diluent quality control samples to determine relative accuracy, intra- and inter-assay precision, limit of detection (LOD), lower limit of quantification (LLOQ), dilutional linearity, and spike recovery. The optimized assay was used to quantify Aβ1-42 in clinical samples from patients treated with the β-site amyloid precursor protein cleaving enzyme 1 inhibitor LY2886721. The prototype assay measured Aβ1-42 with an LOD of 0.3 pg/ml and an LLOQ of 2.8 pg/ml in plasma, calibrated using an Aβ1-42 peptide standard from Fujirebio. Assay precision was acceptable with intra- and inter-assay coefficients of variation both being ≤10%. Dilutional linearity was demonstrated in sample diluent and immunodepleted human plasma. Analyte spike recovery ranged from 51% to 93% with a mean of 80%. This assay was able to quantify Aβ1-42 in all of the 84 clinical samples tested. A rapid reduction in levels of Aβ1-42 was detected within 1 h after drug treatment, and a dose-dependent decrease of Aβ1-42 levels was also observed over the time course of sample collection. This digital ELISA has potential utility in clinical applications for quantification of Aβ1-42 in plasma where high sensitivity and precision are required. |
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Demographic breakdown
Readers by professional status | Count | As % |
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