Department of Instructive Biomaterials Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, 6200 MD Maastricht, The Netherlands.
Mesoscale Chemical Systems Group, MESA+ Institute for Nanotechnology, University of Twente, 7522 NB Enschede, The Netherlands.
Molecules. 2020 Mar 18;25(6):1380. doi: 10.3390/molecules25061380.
A microfluidic protein aggregation device (microPAD) that allows the user to perform a series of protein incubations with various concentrations of two reagents is demonstrated. The microfluidic device consists of 64 incubation chambers to perform individual incubations of the protein at 64 specific conditions. Parallel processes of metering reagents, stepwise concentration gradient generation, and mixing are achieved simultaneously by pneumatic valves. Fibrillation of bovine insulin was selected to test the device. The effect of insulin and sodium chloride (NaCl) concentration on the formation of fibrillar structures was studied by observing the growth rate of partially folded protein, using the fluorescent marker Thioflavin-T. Moreover, dual gradients of different NaCl and hydrochloric acid (HCl) concentrations were formed, to investigate their interactive roles in the formation of insulin fibrils and spherulites. The chip-system provides a bird's eye view on protein aggregation, including an overview of the factors that affect the process and their interactions. This microfluidic platform is potentially useful for rapid analysis of the fibrillation of proteins associated with many misfolding-based diseases, such as quantitative and qualitative studies on amyloid growth.
一种微流控蛋白质聚集装置（microPAD），可使用户对两种试剂的不同浓度进行一系列蛋白质孵育。该微流控装置由 64 个孵育室组成，可在 64 种特定条件下对蛋白质进行单独孵育。气动阀可同时实现试剂的计量、逐步浓度梯度生成和混合的平行过程。选择牛胰岛素的纤颤来测试该装置。通过使用荧光标记物硫黄素-T 观察部分折叠蛋白的生长速率，研究胰岛素和氯化钠（NaCl）浓度对纤维状结构形成的影响。此外，还形成了不同 NaCl 和盐酸（HCl）浓度的双梯度，以研究它们在胰岛素纤维和球晶形成中的相互作用。该芯片系统可全面观察蛋白质聚集，包括影响该过程及其相互作用的因素的概述。这种微流控平台对于与许多错误折叠相关疾病的蛋白质纤颤的快速分析可能非常有用，例如对淀粉样蛋白生长的定量和定性研究。
