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基于具有石墨烯生物界面的石英晶体微天平的即时检测免疫传感器用于抗体检测。

A Point-of-Care Immunosensor Based on a Quartz Crystal Microbalance with Graphene Biointerface for Antibody Assay.

作者信息

Hampitak Piramon, Jowitt Thomas A, Melendrez Daniel, Fresquet Maryline, Hamilton Patrick, Iliut Maria, Nie Kaiwen, Spencer Ben, Lennon Rachel, Vijayaraghavan Aravind

机构信息

Department of Materials and National Graphene Institute, Faculty of Science and Engineering, The University of Manchester, Oxford Road, Manchester M13 9PL, U.K.

School of Biological Sciences, Faculty of Biology Medicine and Health, The University of Manchester, Oxford Road, Manchester M13 9PL, U.K.

出版信息

ACS Sens. 2020 Nov 25;5(11):3520-3532. doi: 10.1021/acssensors.0c01641. Epub 2020 Oct 25.

Abstract

We present a sensitive and low-cost immunoassay, based on a customized open-source quartz crystal microbalance coupled with graphene biointerface sensors (G-QCM), to quantify antibodies in undiluted patient serum. We demonstrate its efficacy for a specific antibody against the phospholipase A2 receptor (anti-PLA2R), which is a biomarker in primary membranous nephropathy. A novel graphene-protein biointerface was constructed by adsorbing a low concentration of denatured bovine serum albumin (dBSA) on the reduced graphene oxide (rGO) sensor surface. The dBSA film prevents the denaturation of the protein receptor on the rGO surface and serves as the cross-linker for immobilization of the receptor for anti-PLA2R antibodies on the surface. The detection limit and selectivity of this G-QCM biosensor was compared with a commercial QCM system. The G-QCM immunoassay exhibited good specificity and high sensitivity toward the target, with an order of magnitude better detection limit (of 100 ng/mL) compared to the commercial system, at a fraction of the cost and with considerable time saving. The results obtained from patient sera compared favorably with those from enzyme-linked immunosorbent assay, validating the feasibility of use in clinical applications. The multifunctional dBSA-rGO platform provides a promising biofunctionalization method for universal immunoassay and biosensors. With the advantages of inexpensive, rapid, and sensitive detection, the G-QCM sensor and instrument form an effective autoimmune disease screening tool.

摘要

我们展示了一种灵敏且低成本的免疫测定方法,该方法基于定制的开源石英晶体微天平与石墨烯生物界面传感器(G-QCM)相结合,用于定量未稀释患者血清中的抗体。我们证明了其对针对磷脂酶A2受体的特异性抗体(抗PLA2R)的有效性,抗PLA2R是原发性膜性肾病的一种生物标志物。通过在还原氧化石墨烯(rGO)传感器表面吸附低浓度的变性牛血清白蛋白(dBSA)构建了一种新型的石墨烯-蛋白质生物界面。dBSA膜可防止rGO表面的蛋白质受体变性,并作为交联剂将抗PLA2R抗体的受体固定在表面。将这种G-QCM生物传感器的检测限和选择性与商业QCM系统进行了比较。G-QCM免疫测定对目标物表现出良好的特异性和高灵敏度,与商业系统相比,检测限低一个数量级(为100 ng/mL),成本仅为其一小部分,且节省大量时间。从患者血清中获得的结果与酶联免疫吸附测定的结果相比具有优势,验证了其在临床应用中的可行性。多功能dBSA-rGO平台为通用免疫测定和生物传感器提供了一种有前景的生物功能化方法。凭借廉价、快速和灵敏检测的优势,G-QCM传感器和仪器形成了一种有效的自身免疫性疾病筛查工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/00a5/7706117/b470ccacab07/se0c01641_0002.jpg

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