Q-DOAS: A Proximity Quenching Assay for Real-Time Detection of Early Protein Aggregation Events
Abstract
Protein aggregation is central to numerous neurodegenerative diseases. The paucity of facile assays to measure aggregation and detect amyloid seeds hinders therapeutics development and clinical assessments. The standard ThT assay reports on amyloid formation but is blind to early oligomers and is prone to interference. We describe Q-DOAS, a simple and reproducible assay that quantifies aggregation in real time via proximity-quenching of a site-specifically conjugated dye (BODIPY-TMR). Using mutant Huntingtin-exon 1 (mHTT-Ex1) and α-Synuclein A53T, we show Q-DOAS detects pre-amyloid oligomers, yielding quantitative kinetic data compatible with mechanistic analysis. We further demonstrate its utility to dissect mutational effects, screen for protein and small-molecule aggregation inhibitors, and quantify amyloid seeding activity in cellular and mouse models of Huntington’s disease and in cerebrospinal fluid from Parkinson’s disease patients without amplification. Q-DOAS provides a sensitive, robust, and scalable tool for studying the earliest events in amyloid pathologies and for advancing therapeutic development.
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BibTeX
@misc{kleczko2026qdoas,
title = {{Q-DOAS: A Proximity Quenching Assay for Real-Time Detection of Early Protein Aggregation Events}},
author = {Korbin Michael Kleczko and Dan Gestaut and Sarah Dobbins and Juliana Abramovich and Cole Sitron and Xia Li and Rebecca Chan and Nan Wang and X. William Yang and F. Ulrich Hartl and Judith Frydman},
howpublished = {bioRxiv},
year = {2026},
month = jun,
doi = {10.64898/2026.06.16.732424},
url = {https://www.biorxiv.org/content/10.64898/2026.06.16.732424v2},
note = {CC0},
}
RIS
TY - UNPB
TI - Q-DOAS: A Proximity Quenching Assay for Real-Time Detection of Early Protein Aggregation Events
AU - Korbin Michael Kleczko
AU - Dan Gestaut
AU - Sarah Dobbins
AU - Juliana Abramovich
AU - Cole Sitron
AU - Xia Li
AU - Rebecca Chan
AU - Nan Wang
AU - X. William Yang
AU - F. Ulrich Hartl
AU - Judith Frydman
PY - 2026
DA - 2026/06/16
PB - bioRxiv
DO - 10.64898/2026.06.16.732424
UR - https://www.biorxiv.org/content/10.64898/2026.06.16.732424v2
AB - Protein aggregation is central to numerous neurodegenerative diseases. The paucity of facile assays to measure aggregation and detect amyloid seeds hinders therapeutics development and clinical assessments. The standard ThT assay reports on amyloid formation but is blind to early oligomers and is prone to interference. We describe Q-DOAS, a simple and reproducible assay that quantifies aggregation in real time via proximity-quenching of a site-specifically conjugated dye (BODIPY-TMR). Using mutant Huntingtin-exon 1 (mHTT-Ex1) and α-Synuclein A53T, we show Q-DOAS detects pre-amyloid oligomers, yielding quantitative kinetic data compatible with mechanistic analysis. We further demonstrate its utility to dissect mutational effects, screen for protein and small-molecule aggregation inhibitors, and quantify amyloid seeding activity in cellular and mouse models of Huntington’s disease and in cerebrospinal fluid from Parkinson’s disease patients without amplification. Q-DOAS provides a sensitive, robust, and scalable tool for studying the earliest events in amyloid pathologies and for advancing therapeutic development.
SN - 2692-8205
ER -