One of the most critical pathological features of Alzheimer’s disease (AD) is the accumulation of β-amyloid (Aβ) peptides that form extracellular senile plaques in the brain. Increasing evidence suggests that small soluble oligomers of Aβ are the toxic form of the peptide and may instigate AD. The Aβ peptide is the amyloid precursor protein (APP) digestion product. β-secretase and γ-secretase proteolysis releases Aβ from the cell. In addition to APP, γ-secretase also cleaves many other type I transmembrane (TM) protein substrates. As a crucial enzyme for Aβ production, γ-secretase is an appealing therapeutic target for AD.Gamma Secretase Activity Assay Cell Line from Innoprot allows the identification of γ-secretase activity inhibitors.

Innoprot performs High Throughput Imaging assays using this model to identify γ-secretase activity inhibitors. Results indicates that the pharmacological inhibition of γ-secretase implicated in AD remains a valid strategy for drug screening. After treatments, we obtain images and analyze them using Attovision software from Becton Dickinson and we quantify fluorescent vesicles into the cytoplasm.

Results indicate that the detecting dynamyc range is dependent on the inhibitor biophysics and biochemical characteristics and the treatment time. This retention assay shows an average of Z´= 0.79+/- 0.02 for High Content Screening with a 24 hours treatment.

Applications

• This model permits evaluate lybraries of compounds, candidates to γ-secretase inhibitors, in living cells studying the vesicles retention
• Gamma Secretase Activity Assay provides a strategy to evaluate drug against γ-secretase activity without the necessity to be permeable.
• This model allows to analyse in the space and time the compund effect in a multiparametric manner.