Single Chain FV fragments as a tool for Aβ oligomers detection

GONZALEZ GARELLO, TOMAS; CERCATO, MAGALI C.; BAEZ M. VERÓNICA; ACUTAIN, M. FLORENCIA; EPSTEIN, ALBERTO L.; JERUSALINSKY, DIANA A.; KORNISIUK, EDGAR; SALVETTI, ANNA

Mar del Plata

Congreso; SAIC. SAI. SAFIS 2018; 2018

Sociedad Argentina de Investigacion Clinica

Alzheimer's disease (AD) is a neurodegenerative disorder of the Central Nervous System that affects millions of people in the world. AD involves progressive lost in cognitive functions due to neuronal death in hippocampus and other related areas. From the Aβ theory point of view, AD was first characterized by the presence of amyloid plaques composed by Aβ peptides aggregates. Years after, it was discovered that neither Aβ peptides, nor amyloid plaques were responsible for synaptic failures and neuronal death. In parallel, it was shown that soluble Aβ aggregates (from 3 to 50 monomers) called Aβ oligomers (AβOs) were the main toxins in this pathology. Currently, although palliative therapy for AD exists, there are no treatments that inhibit the progression of this pathology. The development of new biotechnology tools that help to investigate changes in AβOs levels at AD earlier stages would allow delaying the onset or progression of EA symptoms through secondary prevention therapies, leading to an improvement in AD patient's quality of life. In this context we built an Adeno associated Vector (AAV) for transiently expressing a single chain variable fragment antibody (scFv) that binds specifically AβOs (AAV-scFv-NUSC1Glu). Thus, we sought to develop an assay that allows us to differentially detect AβOs levels in AD models samples. We expressed the AAV-scFv-NUSC1Glu in cell lines and collected the scFv from the supernatant. Then we tested two different ELISA assay models for AβOs detection: A Direct ELISA and a Competitive ELISA. Preliminary results showed that a competition essay was more effective for ABOs detection. However, more essays could be performed in order to improve ABOs detection.