Institut de Ciència de Materials de Barcelona (ICMAB-CSIC), Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Campus de La Universitat Autònoma de Barcelona, Cerdanyola, 08193, Barcelona, Spain.
Institut de Ciència de Materials de Barcelona (ICMAB-CSIC), Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Campus de La Universitat Autònoma de Barcelona, Cerdanyola, 08193, Barcelona, Spain; Department of Chemical Sciences, University of Padua, via Francesco Marzolo 1, 35131, Padova, Italy.
Biosens Bioelectron. 2020 Nov 1;167:112433. doi: 10.1016/j.bios.2020.112433. Epub 2020 Jul 24.
The aggregation of α-synuclein is a critical event in the pathogenesis of neurological diseases, such as Parkinson or Alzheimer. Here, we present a label-free sensor based on an Electrolyte-Gated Organic Field-Effect Transistor (EGOFET) integrated with microfluidics that allows for the detection of amounts of α-synuclein in the range from 0.25 pM to 25 nM. The lower limit of detection (LOD) measures the potential of our integrated device as a tool for prognostics and diagnostics. In our device, the gate electrode is the effective sensing element as it is functionalised with anti-(α-synuclein) antibodies using a dual strategy: i) an amino-terminated self-assembled monolayer activated by glutaraldehyde, and ii) the His-tagged recombinant protein G. In both approaches, comparable sensitivity values were achieved, featuring very low LOD values at the sub-pM level. The microfluidics engineering is central to achieve a controlled functionalisation of the gate electrode and avoid contamination or physisorption on the organic semiconductor. The demonstrated sensing architecture, being a disposable stand-alone chip, can be operated as a point-of-care test, but also it might represent a promising label-free tool to explore in-vitro protein aggregation that takes place during the progression of neurodegenerative illnesses.
α-突触核蛋白的聚集是神经退行性疾病(如帕金森病或阿尔茨海默病)发病机制中的一个关键事件。在这里,我们提出了一种基于无标记传感器的方法,该传感器基于带有微流控系统的电解质门控有机场效应晶体管(EGOFET),可检测 0.25 pM 至 25 nM 范围内的 α-突触核蛋白含量。检测的下限(LOD)衡量了我们集成设备作为预测和诊断工具的潜力。在我们的设备中,栅极电极是有效的传感元件,因为它使用双重策略与抗(α-突触核蛋白)抗体功能化:i)戊二醛激活的末端氨基自组装单层,和 ii)His 标记的重组蛋白 G。在这两种方法中,都实现了相当的灵敏度值,具有非常低的亚皮摩尔级别的 LOD 值。微流控工程对于实现栅极电极的受控功能化至关重要,可以避免污染或物理吸附在有机半导体上。所展示的传感架构是一种一次性的独立芯片,可以作为即时检测进行操作,也可能代表一种有前途的无标记工具,用于探索神经退行性疾病进展过程中发生的体外蛋白质聚集。