Label-free and reagent-less protein biosensing using aptamer-modified extended-gate field-effect transistors
Label-free and reagent-less protein biosensing using aptamer-modified extended-gate field-effect transistors
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DOI：
10.1016/j.bios.2013.01.053
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发表时间：
2013-07-15
影响因子：
12.6
通讯作者：
Miyahara, Yuji
中科院分区：
文献类型：
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作者：
Goda, Tatsuro;Miyahara, Yuji
我们已经开发了基于适配体修饰的场效应晶体管（FET）的生物传感器检测溶菌酶和凝血酶。作为这些模型蛋白的敏感和特异性配体的寡核苷酸适配体共价固定在延伸到FET栅极的金电极上，与巯基分子一起形成密集的自组装单层（SAM）。基于适体的电位法是在一个多平行的方式使用微电极阵列格式的门电极。在缓冲溶液中将各种浓度的靶蛋白引入功能化电极后，实时监测门电位的变化。特异性蛋白质结合改变了门/溶液界面的电荷密度，即，界面电位，因为捕获的蛋白质的固有局部净电荷。该电位法成功地测定了固相上的溶菌酶和凝血酶，其动态范围分别为15.2-1040 nM和13.4-1300 nM，检测限分别为12.0 nM和6.7 nM。重要的是，即使在加标10%胎牛血清（FBS）的条件下，也可通过特异性蛋白质识别获得稳健的信号。这里描述的技术都在互补金属氧化物半导体（CMOS）兼容格式内，并且因此有希望用于高效和低成本的制造，并且借助于先进的半导体处理技术而易于缩小尺寸和集成。(C)2013爱思唯尔有限公司版权所有。
We have developed biosensors based on an aptamer-modified field-effect transistor (FET) for the detection of lysozyme and thrombin. An oligonucleotide aptamer as a sensitive and specific ligand for these model proteins was covalently immobilized on a gold electrode extended to the gate of FET together with thiol molecules to make a densely packed self-assembled monolayer (SAM). The aptamer-based potentiometry was achieved in a multi-parallel way using a microelectrodes array format of the gate electrode. A change in the gate potential was monitored in real-time after introduction of a target protein at various concentrations to the functionalized electrodes in a buffer solution. Specific protein binding altered the charge density at the gate/solution interface, i.e., interface potential, because of the intrinsic local net-charges of the captured protein. The potentiometry successfully determined the lysozyme and thrombin on the solid phase with their dynamic ranges 15.2-1040 nM and 13.4-1300 nM and the limit of detection of 12.0 nM and 6.7 nM, respectively. Importantly, robust signals were obtained by the specific protein recognition even in the spiked 10% fetal bovine serum (FBS) conditions. The technique herein described is all within a complementary metal oxide semiconductor (CMOS) compatible format, and is thus promising for highly efficient and low cost manufacturing with the readiness of downsizing and integration by virtue of advanced semiconductor processing technologies. (C) 2013 Elsevier B.V. All rights reserved.