Jain Shubha, Swain Sarpras, Das Lopamudra, Swain Sarita, Giri Lopamudra, Kondapi Anand Kumar, Narayanan Unni Harikrishnan
Department of Biomedical Engineering, Indian Institute of Technology Hyderabad, Hyderabad 502285, India.
Department of Chemical Engineering, Indian Institute of Technology Hyderabad, Hyderabad 502285, India.
Bioengineering (Basel). 2020 Dec 13;7(4):162. doi: 10.3390/bioengineering7040162.
Tau protein aggregation is identified as one of the key phenomena associated with the onset and progression of Alzheimer's disease. In the present study, we performed on-chip confocal imaging of tau protein aggregation and tau-drug interactions using a spiral-shaped passive micromixing platform. Numerical simulations and experiments were performed in order to validate the performance of the micromixer design. We performed molecular modeling of adenosine triphosphate (ATP)-induced tau aggregation in order to successfully validate the concept of helical tau filament formation. Tau aggregation and native tau restoration were realized using an immunofluorescence antibody assay. The dose-response behavior of an Alzheimer's drug, methylthioninium chloride (MTC), was monitored on-chip for defining the optimum concentration of the drug. The proposed device was tested for reliability and repeatability of on-chip tau imaging. The amount of the tau protein sample used in our experiments was significantly less than the usage for conventional techniques, and the whole protein-drug assay was realized in less than two hours. We identified that intensity-based tau imaging could be used to study Alzheimer's drug response. In addition, it was demonstrated that cell-free, microfluidic tau protein assays could be used as potential on-chip drug evaluation tools for Alzheimer's disease.
tau蛋白聚集被认为是与阿尔茨海默病的发病和进展相关的关键现象之一。在本研究中，我们使用螺旋形被动微混合平台对tau蛋白聚集和tau-药物相互作用进行了芯片上共聚焦成像。进行了数值模拟和实验以验证微混合器设计的性能。我们对三磷酸腺苷（ATP）诱导的tau聚集进行了分子建模，以成功验证螺旋形tau细丝形成的概念。使用免疫荧光抗体测定法实现了tau聚集和天然tau的恢复。在芯片上监测阿尔茨海默病药物亚甲蓝（MTC）的剂量反应行为，以确定药物的最佳浓度。对所提出的装置进行了芯片上tau成像的可靠性和重复性测试。我们实验中使用的tau蛋白样品量明显少于传统技术的用量，并且整个蛋白质-药物测定在不到两小时内完成。我们发现基于强度的tau成像可用于研究阿尔茨海默病药物反应。此外，还证明了无细胞微流控tau蛋白测定可作为阿尔茨海默病潜在的芯片上药物评估工具。
