Alzheimer’s Disease (AD) is characterized by brain depositions of the beta amyloid (Aβ). 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 g-secretase proteolysis releases Aβ from the cell. APP Processing Assay Cell Line from Innoprot allows the identification of Aβ peptide formation inhibitors such as secretase activity inhibitors.

Innoprot performs High Throughput Imaging assays using this model, selecting both γ and β secretase commercial inhibitors as reference compounds. Results indicates that the pharmacological inhibition of secretases implicated in AD remains a valid strategy for drug screening. Therefore, this cellular model allows to monitor the secretases activity in vivo treated with a library compounds in an automatic epifluorescent imaging system to acquire and analyze how robust the model is. 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´= 0.71+/- 0.01 for High Content Screening with a 72 hours treatment.

Applications

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