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P2042
Comparison of An Alphalisa Assay vs. ECL & ELISA Assays to Measure Abeta40 & Abeta42 Levels for HTS & Lead Optimization
Presenter Bethany Hughes, Merck, USA
Additional Authors: Hughes, B.L., Smotrov, N., Zhang, S., Shah, S., Bays, N.W., Kariv, I., Moxham, C.M.
Increased levels of the Amyloid protein A-beta (Aâ) are associated with Alzheimer's Disease (AD), and therefore the ability to quantitate Aâ levels in vitro and in vivo is critical in drug discovery efforts aimed at lowering Aâ levels. While different assay platforms have been used to measure Aâ42 and Aâ40, they are not readily amenable to both HTS and Lead Optimization activities in that they are not easily miniaturized or that they require several wash steps. AlphaLisa is a novel homogenous assay technology which has been shown to be robust and scalable for a variety of analytes. We chose to compare AlphaLisa vs. two other assay formats previously utilized to quantify Aâ42 and Aâ40, namely electrochemiluminescence (ECL) and ELISA. The extent of testing was conducted using purified Aâ peptides, cell supernatants and cerebrospinal fluid (CSF). While all 3 assays were shown to be robust (i.e. Z' >0.5) and capable of distinguishing molecules with different mechanisms of action, AlphaLisa was superior when one takes into account cost/well, amenability to miniaturization, and assay cycle-time.