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用于超高灵敏度和特异性检测生物素的石墨烯场效应晶体管生物传感器。

Graphene field-effect transistor biosensor for detection of biotin with ultrahigh sensitivity and specificity.

作者信息

Wang Shiyu, Hossain Md Zakir, Shinozuka Kazuo, Shimizu Natsuhiko, Kitada Shunya, Suzuki Takaaki, Ichige Ryo, Kuwana Anna, Kobayashi Haruo

机构信息

Division of Electronics and Informatics, Graduate School of Science and Engineering, Gunma University 1-5-1 Tenjin-cho, Kiryu, 376-8515, Japan.

Gunma University Initiative for Advanced Research (GIAR), Gunma University, Japan.

出版信息

Biosens Bioelectron. 2020 Oct 1;165:112363. doi: 10.1016/j.bios.2020.112363. Epub 2020 Jun 4.

Abstract

Because avidin and biotin molecules exhibit the most specific and strongest non-covalent interaction, avidin-biotin technology is widely used in ELISA (enzyme-linked immunosorbent assay) kits for the detection of different bio-macromolecules linked to different diseases including cancer and influenza. Combining the outstanding electrical conductivity (200,000 cmVs) of graphene with the unique avidin and biotin interaction, we demonstrate a novel graphene field-effect transistor (GFET) biosensor for the quantitative detection of bio-macromolecules. The GFET consists of six pairs of interdigital Cr/Au electrodes supported on Si/SiO substrate with an avidin immobilized single layer graphene channel as the sensing platform. By monitoring the real time current change upon the addition of biotin solution in bovine serum albumin (BSA) in the silicone pool preformed onto the GFET, the lowest detectable biotin concentration is estimated to be 90 fg/ml (0.37 pM). The specificity of the GFET is confirmed both by controlled and real sample measurements. From the magnitude of current change upon the addition of different concentrations of biotin solutions, the dissociation constant K is estimated to be 1.6 × 10 M. Since biotin is capable of conjugating with proteins, nucleotides and other bio-macromolecules without altering their properties, the present GFET sensor with its ultra-high sensitivity (0.37 pM) and specificity can be tailored to the rapid point-of-care detection of different types of desired biomolecules at very low concentration level through biotinylation as well as the exogenous biotin in blood serum.

摘要

由于抗生物素蛋白和生物素分子表现出最特异且最强的非共价相互作用,抗生物素蛋白-生物素技术被广泛应用于酶联免疫吸附测定(ELISA)试剂盒中,用于检测与包括癌症和流感在内的不同疾病相关的各种生物大分子。将石墨烯出色的电导率(200,000 cmVs)与独特的抗生物素蛋白和生物素相互作用相结合,我们展示了一种用于定量检测生物大分子的新型石墨烯场效应晶体管(GFET)生物传感器。该GFET由支撑在Si/SiO衬底上的六对叉指式Cr/Au电极组成,以固定有抗生物素蛋白的单层石墨烯通道作为传感平台。通过监测在预先形成于GFET上的硅树脂池中向牛血清白蛋白(BSA)中添加生物素溶液时的实时电流变化,估计可检测到的最低生物素浓度为90 fg/ml(0.37 pM)。通过对照测量和实际样品测量均证实了GFET的特异性。根据添加不同浓度生物素溶液时电流变化的幅度,估计解离常数K为1.6×10⁻⁸ M。由于生物素能够与蛋白质、核苷酸和其他生物大分子结合而不改变它们的性质,因此本GFET传感器凭借其超高灵敏度(0.37 pM)和特异性,可通过生物素化以及血清中的外源性生物素,针对在非常低浓度水平下快速即时检测不同类型的所需生物分子进行定制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/74fb/7272179/601d282aa975/gr1_lrg.jpg

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