Performance of novel tau antibodies across multiple modalities for Alzheimer's disease assessment.

INTRODUCTION: Epitope-specific antibodies are crucial for accurately identifying pathologic forms of proteins like hyperphosphorylated tau in Alzheimer's disease (AD). METHODS: We developed monoclonal antibodies targeting phosphorylated tau (p-tau) at sites pT181, pT217, and pT231 using peptide immunogens. Antibody performance was assessed using peptide immunoprecipitation-mass spectrometry, western blot, and quantitative immunohistochemistry in brain, and single-molecule array immunoassay in cerebrospinal fluid (CSF). RESULTS: These antibodies demonstrated specific immunoprecipitation of phosphorylated peptides in phosphate-buffered saline, CSF, and plasma. Quantitative immunostaining in brain tissue highlighted neuronal tau pathology and were negative in control tissues (P < 0.01 for mice; P < 0.001 for humans). Western blotting with recombinant p-tau and post mortem AD brain tissue samples provided further validation. In a pilot CSF immunoassay these antibodies performed similarly to commercial antibodies (R(2 )> 0.997) and similarly differentiated probable AD from controls (P < 0.01). DISCUSSION: Overall, our work contributes new reagents for AD research, specifically anti-p-tau antibodies, for the scientific community. HIGHLIGHTS: Novel phosphorylated tau pT181, pT217, and pT231 antibodies are sensitive and specific. The antibodies perform well across multiple modalities for diagnosis and research. The cerebrospinal fluid assay with these antibodies is similar to a commercial assay (Quanterix). The antibodies are freely accessible from the Developmental Studies Hybridoma Bank.