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bioRxiv · 10.64898/2026.01.07.698009

Protocol for using an ELISA assay to detect total α-synuclein levels in Drosophila melanogaster lines expressing human α-synuclein point mutations

Sciortino, M.·Velazquez, R.·Tillmon, H.·Banerjee, S.
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Abstract

In Parkinsons disease (PD), aggregation of alpha-synuclein (-syn) contributes to neuronal dysfunction and death, particularly in dopaminergic neurons, driving disease progression. Several pathogenic point mutations have been identified in human -syn such as A30P, E46K, H50Q, G51D, A53T and A53E that have been associated with PD. In this study, a sandwich ELISA assay was developed to quantify total -syn levels in various Drosophila melanogaster genotypes expressing A30P, E46K, G51D and A53T. Using this approach, we compared -syn levels across wild-type (WT) and mutant forms of the protein. The E46K and A53T mutations exhibited higher total -syn concentrations compared to WT and the G51D mutation. In addition to characterizing mutationdependent differences in -syn levels, this assay was applied to evaluate smallmolecule modulators that inhibit -syn aggregation. These findings demonstrate that the ELISA-based approach provides a useful platform for quantifying -syn and assessing potential therapeutic compounds targeting syn pathology.

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Sciortino, M., Velazquez, R., Tillmon, H., Banerjee, S.. 2026-01-07. Protocol for using an ELISA assay to detect total α-synuclein levels in Drosophila melanogaster lines expressing human α-synuclein point mutations. https://doi.org/10.64898/2026.01.07.698009

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