Assays for quantifying aggregated and phosphorylated (S129) human α-synuclein protein are widely used to evaluate pathological burden in patients suffering from synucleinopathy disorders. Many of these assays, however, do not cross-react with mouse α-synuclein or exhibit poor sensitivity for this target, which is problematic considering the preponderance of mouse models at the forefront of pre-clinical α-synuclein research. In this project, we addressed this unmet need by reformulating two existing AlphaLISA^® SureFire^® Ultra™ total and pS129 α-synuclein assay kits to yield robust and ultrasensitive (LLoQ ≤ 0.5 pg/mL) quantification of mouse and human wild-type and pS129 α-synuclein protein. We then employed these assays, together with the BioLegend α-synuclein aggregate ELISA, to assess α-synuclein S129 phosphorylation and aggregation in different mouse brain tissue preparations. Overall, we highlight the compatibility of these new immunoassays with rodent models and demonstrate their potential to advance knowledge surrounding α-synuclein phosphorylation and aggregation in synucleinopathies.

用于定量聚集和磷酸化 （S129） 人 α-突触核蛋白的测定法广泛用于评估突触核蛋白病患者的病理负担。然而，其中许多检测不与小鼠 α-突触核蛋白发生交叉反应，或者对该靶标的敏感性较差，考虑到处于临床前 α-突触核蛋白研究前沿的小鼠模型的优势，这是有问题的。在本项目中，我们通过重新配制两种现有的 AlphaLISA^® SureFire^® Ultra™ 总和 pS129 α-突触核蛋白检测试剂盒来解决这一未满足的需求，以产生小鼠和人野生型以及 pS129 α-突触核蛋白的稳定且超灵敏 （LLoQ ≤ 0.5 pg/mL） 定量。然后，我们使用这些测定法以及 BioLegend α-突触核蛋白聚集体 ELISA 来评估不同小鼠脑组织制剂中 α-突触核蛋白 S129 磷酸化和聚集。总体而言，我们强调了这些新的免疫测定与啮齿动物模型的兼容性，并证明了它们推进围绕突触核蛋白病中 α-突触核蛋白磷酸化和聚集的认识的潜力。