Alzheimer's disease (AD) is a progressive irreversible neurodegenerative disease, where memory loss and other symptoms are associated with

changes in the brain involving amyloid beta (Aβ) plaques and neurofibrillary tangles of tau protein. Current diagnostic methods are invasive and are

only used to confirm the diagnosis after the onset of clinical symptoms. The aim of this project is to develop a method that will allow the detection

of AD biomarkers in a plasma sample using only a simple and accessible instrumental technique. The latter will be based on a small fluorescent

molecular probe which, when added to plasma, will identify AD biomarkers. Upon binding the probe will show a characteristic fluorescence emission

that can be easily measured. The biomarkers that will be targeted in this research are aggregates of Aβ and tau protein (Tau), which are known to be

present in the blood and reflect pathology of the brain. In designing such probes, we will be the first to use computer-assisted tools, a combination

of machine learning, artificial intelligence, molecular modelling and quantum mechanical methods such as (TD)-DFT. In the search for a hit

compound, we will use in vitro assays to examine the selectivity and binding affinity of the synthesised molecules towards Aβ and Tau aggregates.

Studies in a more complex environment, more similar to a real sample, will be performed using in cellulo assays. A proof-of-concept will also be the

ex vivo testing of hit compounds on post mortem brain tissues obtained from AD patients. Increasing the sensitivity of the method as the

concentrations of Aβ and Tau aggregates in the blood are low will be achieved by binding the hit compounds to magnetic nanobeads, which will

allow the biomarkers to be concentrated in the plasma by a magnet. In addition, we will implement gold nanorods to amplify the fluorescence and

thus the diagnostic signal. Quantitative analysis of AD biomarkers in plasma using the developed fluorescent molecular probe will be performed

using the FIDA ("flow-induced dispersion analysis") method. To achieve all these objectives, the project involves ten research institutions, which will

contribute with their diverse expertise and experience for the realisation of the tasks. The Slovenian members of the project are i) University of

Ljubljana (UL) Faculty of Chemistry and Chemical Technology (PI Dr. Jerneja Kladnik), ii) UL Faculty of Pharmacy (Assist. Prof. Dr. Damijan Knez), iii)

UL Faculty of Medicine (Prof. Dr. Mara Bresjanac), iv) UL Faculty of Computer and Information Science (Assist. Prof. Dr. Matevž Pesek), and v)

"Jožef Stefan" Institute (Assist. Prof. Dr. Slavko Kralj). In addition, research institutions from Europe and the USA are also involved in the project, i.e.

vi) CIC biomaGUNE (Dr. Jordi Llop), vii) David Geffen School of Medicine - UCLA (Prof. DDr. Jorge R. Barrio), viii) Budapest University of Technology

and Economics (Assoc. Prof. Dr. Julianna Oláh), ix) University of Barcelona (Prof. Dr. Raimon Sabaté) and Utrecht University (Prof. Dr. Stefan

Rüdiger).