Department of Materials Engineering, School of Engineering, The University of Tokyo , 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Department of Dermatology, Division of Molecular Medical Science, Graduate School of Biomedical Science, Hiroshima University , 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8551, Japan.
Anal Chem. 2017 Dec 5;89(23):12918-12923. doi: 10.1021/acs.analchem.7b03688. Epub 2017 Nov 15.
In this study, we report the label-free and reliable detection of allergic response using a cell-coupled gate field-effect transistor (cell-based FET). Rat basophilic leukemia (RBL-2H3) cells were cultured as a signal transduction interface to induce allergic reaction on the gate oxide surface of the FET, because IgE antibodies, which bind to Fcε receptors at the RBL-2H3 cell membrane, are specifically cross-linked by allergens, resulting in the allergic response of RBL-2H3 cells. In fact, the surface potential at the FET gate decreased owing to secretions such as histamine from the IgE-bound RBL-2H3 cells, which reacted with the allergen. This is because histamine, as one of the candidate secretions, shows basicity, resulting in a change in pH around the cell/gate interface. That is, the RBL-2H3-cell-based FET used in this study was originally from an ion-sensitive FET (ISFET), whose oxide surface (TaO) with hydroxyl groups is fully responsive to pH on the basis of the equilibrium reaction. The allergic response of RBL-2H3 cells on the gate was also confirmed by estimating the amount of β-hexosaminidase released together with histamine and was analyzed using the electrical properties based on an inflammatory response of secreted histamine with the vascular endothelial cell-based FET. Thus, the allergic responses were monitored in a nonoptical and real-time manner using the cell-based FETs with the cellular layers on the gate, which reproduced the in vivo system and were useful for the reliable detection of the allergic reaction.
在这项研究中，我们报告了使用细胞偶联栅场效应晶体管（基于细胞的 FET）进行无标记和可靠的过敏反应检测。将大鼠嗜碱性白血病（RBL-2H3）细胞培养为信号转导接口，以在 FET 的栅氧化层表面上诱导过敏反应，因为 IgE 抗体与 RBL-2H3 细胞膜上的 Fcε 受体结合，过敏原特异性交联，导致 RBL-2H3 细胞发生过敏反应。事实上，由于 IgE 结合的 RBL-2H3 细胞分泌的组胺等物质与过敏原反应，FET 栅极的表面电势降低。这是因为组胺作为候选分泌物之一具有碱性，导致细胞/栅极界面周围的 pH 值发生变化。也就是说，本研究中使用的基于 RBL-2H3 细胞的 FET 最初是一种离子敏感 FET（ISFET），其具有羟基的氧化层（TaO）基于平衡反应对 pH 完全敏感。通过估计与组胺一起释放的β-己糖胺酶的量来确认 RBL-2H3 细胞在栅极上的过敏反应，并使用基于血管内皮细胞 FET 的分泌组胺的炎症反应的电特性进行分析。因此，使用带有栅极上细胞层的基于细胞的 FET 以非光学和实时方式监测过敏反应，该 FET 再现了体内系统，并且对可靠检测过敏反应很有用。
