Format

Send to

Choose Destination
See comment in PubMed Commons below
Anal Biochem. 2014 Aug 15;459:24-30. doi: 10.1016/j.ab.2014.05.007. Epub 2014 May 20.

AlphaLISA-based high-throughput screening assay to measure levels of soluble amyloid precursor protein α.

Author information

1
Torrey Pines Institute for Molecular Studies, 11350 SW Village Parkway, Port Saint Lucie, 34987 FL, USA.
2
Torrey Pines Institute for Molecular Studies, 11350 SW Village Parkway, Port Saint Lucie, 34987 FL, USA. Electronic address: dminond@tpims.org.

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.

KEYWORDS:

Alzheimer’s disease; Aβ42; CHO cells; HTS assay; NT2 cells; Soluble APPα

PMID:
24857774
DOI:
10.1016/j.ab.2014.05.007
[Indexed for MEDLINE]
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science
    Loading ...
    Support Center