Identification of the Tau aggresome in progressive supranuclear palsy by in situ biotinylation by antibody recognition

Here, we used an unbiased proteomic approach, which combines antibody-mediated proximity-detection of co-aggregated proteins together with mass spectrometry. Biotinylation by antibody recognition (BAR) is a recently developed method, by which a primary antibody recognises the target of interest in fixed samples. A secondary antibody conjugated to horseradish peroxidase (HRP) recognises the primary antibody, and with the addition of biotin phenol and hydrogen peroxide, facilitates biotinylation of proteins in close proximity. Biotinylated proteins are then biochemically isolated and identified using mass spectrometry (MS).

本研究采用无偏蛋白质组学方法，将抗体介导的共聚集蛋白邻近检测技术与质谱（mass spectrometry）相结合。抗体识别生物素化法（BAR，Biotinylation by antibody recognition）是一种新近开发的技术，可在固定样本中通过一抗识别目标靶蛋白。辣根过氧化物酶（HRP，horseradish peroxidase）标记的二抗可识别该一抗，在添加生物素酚与过氧化氢后，可对邻近蛋白质进行生物素标记。随后通过生化分离获取生物素化蛋白，并利用质谱（MS）对其进行鉴定。