Q-DOAS: A Proximity Quenching Assay for Real-Time Detection of Early Protein Aggregation Events

Authors

Kleczko, K. M., Gestaut, D., Dobbins, S., Abramovich, J., Sitron, C. S., Li, L., Chan, R., Wang, N., Yang, X. W., Hartl, F.-U., Frydman, J.

Abstract

Accurate measurement of protein aggregation is essential for studying neurodegenerative diseases. The standard ThT assay reports on amyloid formation but is blind to early oligomers and is prone to interference. We describe Q-DOAS, a plate-reader assay that quantifies protein self-assembly in real time via proximity-quenching of a single, site-specifically conjugated dye (BODIPY-TMR). Using mutant Huntingtin-exon 1 (mHTT-Ex1) and -Synuclein A53T, we show Q-DOAS detects pre-amyloid oligomers, yielding quantitative kinetic data compatible with mechanistic analysis. We demonstrate its utility to dissect mutational effects, screen for protein and small-molecule inhibitors, and quantify amyloid seeding activity in cellular and mouse models of Huntington's disease. Q-DOAS also detects seeds 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.

Preprint server: bioRxiv
The authors list and abstract were imported from bioRxiv on 18 Jun 2026.

Stats

1-terrible, 9-excellent. How would you rate this preprint? Sign in in to submit your rating.

Post a comment

You need to be signed in to post comments. You can sign in here.

Comments

There are no comments yet.