National Center for Advancing Translational Sciences, NIH, 9800 Medical Center Drive, MSC: 3370, Bethesda, MD 20892-3370, USA.
Curr Alzheimer Res. 2013 Sep;10(7):679-87. doi: 10.2174/15672050113109990143.
The microtubule-associated protein (MAP) tau has been implicated in the pathology of numerous neurodegenerative diseases. In the past decade, the hyperphosphorylated and aggregated states of tau protein have been important targets in the drug discovery field for the potential treatment of Alzheimer's disease. Although several compounds have been reported to reduce the hyperphosphorylated state of tau or impact the stabilization of tau, their therapeutic activities are remain to be validated. Recently, reduction of total cellular tau protein has emerged as an alternate intervention point for drug development and a potential treatment of tauopathies. We have developed and optimized homogenous assays, using the AlphaLISA and HTRF assay technologies, for the quantification of total cellular tau protein levels in the SH-SY5Y neuroblastoma cell line. The signal-to-basal ratios were 375 and 5.3, and the Z' factors were 0.67 and 0.60 for the AlphaLISA and HTRF tau assays, respectively. The clear advantages of these homogeneous tau assays over conventional total tau assays, such as ELISA and Western blot, are the elimination of plate wash steps and miniaturization of the assay into 1536-well plate format for the ultra-high-throughput screening of large compound libraries.
微管相关蛋白（MAP）tau 已被牵涉到许多神经退行性疾病的病理学中。在过去的十年中，tau 蛋白的过度磷酸化和聚集状态已成为药物发现领域的重要靶点，有望用于治疗阿尔茨海默病。尽管已经有几种化合物被报道可以降低 tau 的过度磷酸化状态或影响 tau 的稳定，但它们的治疗活性仍有待验证。最近，降低总细胞 tau 蛋白已成为药物开发的另一个干预点，并可能成为 tau 病的治疗方法。我们已经开发并优化了使用 AlphaLISA 和 HTRF 测定技术的均相测定法，用于定量 SH-SY5Y 神经母细胞瘤细胞系中的总细胞 tau 蛋白水平。对于 AlphaLISA 和 HTRF tau 测定法，信号与基础比率分别为 375 和 5.3，Z' 因子分别为 0.67 和 0.60。与传统的总 tau 测定法（如 ELISA 和 Western blot）相比，这些均相 tau 测定法具有明显的优势，例如消除了平板洗涤步骤，并将测定法小型化为 1536 孔板格式，用于高通量筛选大型化合物文库。
