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Detection and quantification of tau aggregation using a membrane filter assay.

Edward Chang1, Jeff Kuret

  • 1Integrated Biomedical Sciences Graduate Program, Department of Molecular and Cellular Biochemistry, The Ohio State University College of Medicine, Columbus, OH 43210, USA.

Analytical Biochemistry
|October 24, 2007
PubMed
Summary

This study evaluates filter trap assays for quantifying tau aggregation, a marker for Alzheimer's disease. Nitrocellulose membranes offer the best sensitivity, enabling accurate estimation of tau aggregation kinetics.

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Area of Science:

  • Neuroscience
  • Biochemistry
  • Molecular Biology

Background:

  • Tau protein aggregation is central to Alzheimer's disease pathogenesis.
  • Neurofibrillary lesions formed by tau are key disease markers.
  • Filter trap assays are used to measure tau aggregation, but their performance is not well-characterized.

Purpose of the Study:

  • To evaluate the performance of filter trap assays in quantifying purified recombinant full-length tau protein aggregation in vitro.
  • To investigate the impact of membrane chemistry on assay sensitivity and accuracy.
  • To compare filter trap assay results with other established methods like electron microscopy.

Main Methods:

  • A 96-well format filter trap assay was optimized using various membrane chemistries (nitrocellulose, polyvinylidene fluoride, cellulose acetate) and porosities.

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  • Purified recombinant full-length tau protein was used for aggregation studies.
  • Assay sensitivity and linearity were assessed across different conditions.
  • Results were compared with electron microscopy measurements.

    • Main Results:

    • Nitrocellulose membranes demonstrated superior assay sensitivity compared to polyvinylidene fluoride and cellulose acetate.
    • Nonlinear correlations were observed between signal intensity and tau concentration across all tested filter conditions.
    • After correcting for nonlinearity, the filter trap assay yielded a critical monomer concentration for tau aggregation that was statistically consistent with electron microscopy findings.

    Conclusions:

    • Filter trap assays can be effectively utilized to quantify tau aggregation kinetics.
    • Optimizing membrane chemistry, particularly using nitrocellulose, enhances assay sensitivity.
    • The filter trap assay, when properly corrected, provides reliable measurements comparable to electron microscopy for assessing tau aggregation.