An optimized filter trap assay for detecting recombinant authentic tau fibrils.

Area of Science:

• Neuroscience
• Biochemistry
• Molecular Biology

Background:

• Tauopathies are neurodegenerative diseases characterized by the aggregation of tau protein.
• Existing methods often use non-physiological tau fibrils, limiting disease relevance.
• Authentic tau fibrils can be formed by a truncated tau fragment (dGAE) without polyanions.

Purpose of the Study:

• To develop and optimize a sensitive Filter Trap Assay (FTA) for detecting authentic tau fibrils in vitro.
• To establish a physiologically relevant method for tau fibril detection.
• To evaluate the efficacy of sarkosyl in preserving tau fibril integrity.

Main Methods:

• Optimization of the Filter Trap Assay (FTA) using nitrocellulose membranes.
• Utilizing the truncated tau fragment (297-391), also termed dGAE, to form authentic tau fibrils.
• Employing sarkosyl for protein lysate preparation and preservation of tau fibril aggregates.

Main Results:

• The refined FTA demonstrates heightened sensitivity in detecting authentic tau fibrils.
• Sarkosyl proved superior in preserving the aggregated state of tau dGAE fibrils in vitro.
• The assay provides a user-friendly and cost-effective method for tau fibril detection.

Conclusions:

• The optimized FTA offers a reliable method for studying authentic tau fibrils in a relevant context.
• This advancement facilitates broader research into tauopathies and neurodegenerative disorders.
• The methodology bridges basic research with advanced structural analysis of tau aggregates.