Open AccessDevelopment of Homogeneous 384-Well High-Throughput Screening Assays for Aβ1-40 and Aβ1-42 Using AlphaScreen™ Technology
Author(s) -
Philip G. Szekeres,
Kaitlin Leong,
Theresa A. Day,
Ann E. Kingston,
Eric Karran
Publication year - 2008
Publication title -
slas discovery
Language(s) - English
Resource type - Journals
eISSN - 2472-5560
pISSN - 2472-5552
DOI - 10.1177/1087057107312778
Subject(s) - homogeneous , high throughput screening , computational biology , amyloid beta , computer science , amyloid (mycology) , chemistry , chromatography , peptide , biochemistry , biology , mathematics , combinatorics , inorganic chemistry
Amyloid beta (Abeta) peptides are the major constituent of amyloid plaques, one of the hallmark pathologies of Alzheimer's disease. Accurate and precise quantitation of these peptides in biological fluids is a critical component of Alzheimer's disease research. The current most established assay for analysis of Abeta peptides in preclinical research is enzyme-linked immunosorbent assay (ELISA), which, although sensitive and of proven utility, is a multistep, labor-intensive assay that is difficult to automate completely. To overcome these limitations, the authors have developed and optimized simple, sensitive, homogeneous 384-well assays for Abeta1-42 and Abeta1-40 using AlphaScreen technology. The assays are capable of detecting Abeta peptides at concentrations <2 pg/mL and, using a final assay volume of 20 microL, routinely generate Z' values >0.85. The AlphaScreen format has the following key advantages: substantially less hands-on time to run, easier to automate, higher throughput, and less expensive to run than the traditional ELISA. The results presented here show that AlphaScreen technology permits robust, efficient, and cost-effective quantitation of Abeta peptides.