title: Detection and quantification of tau aggregation using a membrane filter assay.
abstract: Aggregation of the microtubule-associated protein tau contributes to the formation of neurofibrillary lesions in Alzheimer's disease and is a useful marker of disease progression. Although filter trap assays have been employed to assess the extent of tau aggregation in cells and tissues as well as in vitro, their performance relative to other assay modalities has not been reported. To clarify this issue, the ability of the filter trap approach to quantify aggregation of purified recombinant full-length tau protein in vitro was examined as a function of membrane chemistry in a 96-well format. Results showed that nitrocellulose yielded the greatest assay sensitivity relative to polyvinylidene fluoride or cellulose acetate at equal membrane porosity. However, all combinations of filter chemistries, porosities, and monoclonal detection antibodies yielded nonlinear correlations between signal intensity and analyte concentration. When corrected for nonlinearity, the filter trap assay determined a value for the critical monomer concentration for tau aggregation that was statistically identical to determinations made by electron microscopy assay. The data suggest conditions under which filter trap assays can be used to estimate tau aggregation kinetics.
authors.affiliation: Integrated Biomedical Sciences Graduate Program, Department of Molecular and Cellular Biochemistry, The Ohio State University College of Medicine, Columbus, OH 43210, USA.
authors.forename: Edward
authors.initials: E
authors.lastname: Chang
authors.forename: Jeff
authors.initials: J
authors.lastname: Kuret
mesh_terms: D000911:Antibodies, Monoclonal; D005374:Filtration / Q000379:methods*; D006801:Humans; D008567:Membranes, Artificial; D008854:Microscopy, Electron; D020836:Protein Structure, Quaternary; D016875:tau Proteins / Q000032:analysis* / Q000737:chemistry* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; C054369:MAPT protein, human; D008567:Membranes, Artificial; D016875:tau Proteins