Fully synthetic phosphorylated Tau181, Tau217, and Tau231 calibrators for Alzheimer’s disease diagnosis

• DOI: 10.3389/fnagi.2023.1340706
• Publication Date: 2024-01-15T04:20:02Z
• Authors: Xinyu Li; Huimei Zeng; Pradeepraj Durairaj; Weihuan Wen; Tianpeng Li; Yanru Zhao; Yang Liu; Xue Liu; Lingpeng Zhan; Lang Rao; Wen Yuan; Tengfei Guo; Weijun Shen; Hui Cai; Zhicheng Chen
• Keywords: Simoa; 03 medical and health sciences; 0302 clinical medicine; diagnosis; calibrator; Aging Neuroscience; Neurosciences. Biological psychiatry. Neuropsychiatry; immunoassay; Tau; Alzheimer’s disease; RC321-571

Background The calibrator in immunoassay plays an essential role diagnosing Alzheimer’s disease (AD). Presently, the most well-studied biomarkers for AD diagnosis are three phosphorylated Tau (p-Tau): p-Tau231, p-Tau217, and p-Tau181. Glycogen synthase-3beta (GSK3β)-phosphorated Tau-441 is commonly used p-Tau immunoassays. However, batch-to-batch inconsistency issue of GSK3β-phosphorylated limits its clinical application. Methods We have successfully generated characterized 61 monoclonal antibodies (mAbs) with distinct epitopes by using hybridoma technique employed them as capture or detection Through chemical synthesis, we synthesized calibrators, which peptides including antibody epitopes, application immunoassays that detect novel calibrators were applied to Enzyme-linked immunosorbent assay (ELISA) Single-molecule array (Simoa) platforms validate their applicability establish a range Results By employing technique, 49 mAbs recognizing (1–22), nine targeting one mAb two p-Tau181 developed. Peptides, recognition antibodies, synthesized. These develop 60 on ELISA platform, six highly sensitive selected ultra-sensitive Simoa platform. Remarkably, LODs 2.5, 2.4, 31.1, 32.9, 46.9, 52.1 pg/ml, respectively. Conclusion Three validated, solved Tau-441. exhibit potential promote standardization diagnostic calibrators. Furthermore, established series specific platform based moved steady step forward diagnosis.