AB Oligomers Detection by a Specific scFv Codified in an AAV Vector

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

Cordoba

Congreso; XXXIII Reunión Anual de la Sociedad Argentina de Investigacion en Neurociencias; 2018

Sociedad Argentina de Investigacion en Neurociencias

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. AD was first characterized by the presence of amyloid plaques composed by Ab peptides aggregates. Although it has been shown that neither Ab peptides nor amyloid plaques were directly responsible for synaptic failures and neuronal death, it was suggested that soluble Ab aggregates (from 4 to 50 monomers), the Ab oligomers (AbOs), were the main toxins at early steps of this pathology. Moreover, elevated AbOs levels have been reported in AD rat models, even before neurodegeneration signs appear. In this context, we built an Adeno Associated Vector (AAV) for transiently expressing a single-chain variable fragment antibody (scFv) that specifically binds AbOs and bears a signal peptide to be secreted (AAV-scFv-NUSC1Glu). N2A y B104 cell lines were infected with AAV-scFv-NUSC1Glu, and the supernatant was then collected. First, we checked scFv expression at mRNA level, by PCR, and protein level by Western blot. Then, we attempted to detect synthetic AbOs levels by ELISA essays. We designed two different essays: A Direct ELISA and a Competitive one. Preliminary results have shown that only the competition essay was useful to discriminate the tested AbOs levels.