Neurochemistry and Neuropharmacology
Author: Silvana Estefanía Soliz Santander | Email: tefi.9625@gmail.com
Silvana Estefanía Soliz Santander1°, Ma. Belén Machin1°, Agustín Pernicone3°, Florencia González-Lizarraga1°, Verónica Manzano3°, César Ávila1°, Oscar Varela2°, Rosana Chehín1°, Esteban Vera Pingitore1°
1° IMMCA (CONICET-UNT-SIPROSA). Pje Dorrego 1080. San Miguel de Tucumán. Tucumán
2° CIHIDECAR (CONICET-UBA). Ciudad Universitaria, Pabellón 2. Ciudad Autónoma de Buenos Aires
Detecting amyloid-like α-synuclein (aSyn) aggregated species in biological fluids is crucial for early Parkinson’s disease (PD) diagnosis and the effectiveness of therapies. However, current diagnostics face challenges because traditional antibodies struggle to detect the wide variety of toxic aSyn species. Based on our previous research demonstrating that certain tetracycline derivatives selectively bind to aggregated α-Syn but not to its monomeric form, we immobilized a modified tetracycline and tested its ability to distinguish between aggregated and monomeric aSyn species using a novel immunoassay without any antibody as a capture molecule. The analyte (aggregated aSyn) was produced from human recombinant aSyn expressed and purified in our laboratory and then aggregated following standard protocols. Our protocol demonstrated nanomolar affinity for aggregated α-synuclein species, with no cross-reactivity to native α-synuclein, which served as a negative control. We also tested this novel ELISA in real cerebrospinal fluid (CSF) samples to detect possible interferents and design strategies to address these issues. The successful application of this novel tetracycline-based immunoassay in real CSF samples highlights its potential for early and more accurate detection of PD’s main biomarker. This advancement not only contributes to more reliable PD diagnostics but also paves the way for further research and development in neurodegenerative disease detection and management.