Investigating Prions and Protein Aggregation at High Sensitivity and Throughput
With no cure available for prion diseases, researchers are striving to improve assay sensitivity and exploit the benefits of real-time monitoring to support the development of better diagnostic and treatment approaches. The post Investigating Prions and Protein Aggregation at High Sensitivity and Throughput appeared first on GEN - Genetic Engineering and Biotechnology News .
Protein misfolding, a key factor in various neurological and non-neurological diseases, has garnered increasing attention from researchers. One such disease, prion disease, is caused by misfolded proteins that accumulate in the nervous system, leading to neurodegeneration and death. Detecting these misfolded proteins, known as prions, is crucial for understanding and developing treatments for these diseases.
Researchers have developed the RT-QuIC (Real-time quaking-induced conversion) assay, a highly sensitive and amplifying assay that detects disease-associated prion activity in real-time. This assay measures the formation of amyloid proteins using the fluorescent dye thioflavin T (ThT), which binds to beta-sheet-rich amyloid structures and increases its emission upon binding.
The RT-QuIC assay generates characteristic amplification curves with a baseline phase, an exponential increase in fluorescence, and a plateau phase. These curves provide quantitative or semi-quantitative measurements of prion seeding activity. Microplate readers, which combine sensitive kinetic fluorescence measurements with periodic cycles of rigorous shaking and temperature control, are the most widely adopted systems for RT-QuIC assays.
The FLUOstar ® Omega microplate reader from BMG LABTECH is a popular choice, offering extended shaking periods and consistent temperature control. Beyond prion diseases, protein misfolding is implicated in a wide range of conditions, including Alzheimer's, Parkinson's, Huntington's, and amyotrophic lateral sclerosis. BMG LABTECH's application notes demonstrate the use of ThT assays to measure the accumulation of misfolded amyloid-beta protein aggregates linked to Alzheimer's disease.
The incorporation of ThT into newly formed amyloid-beta fibrils can be tracked over time, with the formation plateau marking the endpoint of amyloid formation. These ThT assays offer applications across multiple disease areas and research fields, highlighting the importance of understanding protein misfolding and aggregation in various biological processes.
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