title: AlphaLISA-based high-throughput screening assay to measure levels of soluble amyloid precursor protein α.
abstract: Activation of nonamyloidogenic processing of amyloid precursor protein (APP) has been hypothesized to be a viable approach for Alzheimer's disease drug discovery. However, until recently, the lack of HTS-compatible assay technologies precluded large scale screening efforts to discover molecules that potentiate nonamyloidogenic pathways. We have developed an HTS-compatible assay based on AlphaLISA technology that quantitatively detects soluble APPα (sAPPα), a marker of nonamyloidogenic processing of APP, released from live cells in low volume, 384-well plates. The assay exhibited good QC parameters (Z'>0.5, S/B>2). A pilot screen of 801 compounds yielded a novel chemotype that increased the release of sAPPα 2-fold at 5μM. These results suggest that the AlphaLISA-based HTS assay is robust and sensitive and can be used to screen large compound collections to discover molecules that potentiate the release of sAPPα. Additionally, we demonstrated that increase of APP processing by nonamyloidogenic pathways will result in decrease of release of amyloidogenic Aβ40 fragments.
authors.affiliation: Torrey Pines Institute for Molecular Studies, 11350 SW Village Parkway, Port Saint Lucie, 34987 FL, USA.
authors.forename: Hongjie
authors.initials: H
authors.lastname: Wang
authors.affiliation: Torrey Pines Institute for Molecular Studies, 11350 SW Village Parkway, Port Saint Lucie, 34987 FL, USA.
authors.forename: Adel
authors.initials: A
authors.lastname: Nefzi
authors.affiliation: Torrey Pines Institute for Molecular Studies, 11350 SW Village Parkway, Port Saint Lucie, 34987 FL, USA.
authors.forename: Gregg B
authors.initials: GB
authors.lastname: Fields
authors.affiliation: Torrey Pines Institute for Molecular Studies, 11350 SW Village Parkway, Port Saint Lucie, 34987 FL, USA.
authors.forename: Madepalli K
authors.initials: MK
authors.lastname: Lakshmana
authors.affiliation: Torrey Pines Institute for Molecular Studies, 11350 SW Village Parkway, Port Saint Lucie, 34987 FL, USA. Electronic address: dminond@tpims.org.
authors.forename: Dmitriy
authors.initials: D
authors.lastname: Minond
mesh_terms: D016564:Amyloid beta-Protein Precursor / Q000737:chemistry* / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D012995:Solubility
chemical_list: D016564:Amyloid beta-Protein Precursor
keywords: Alzheimer’s disease; Aβ42; CHO cells; HTS assay; NT2 cells; Soluble APPα