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使用通过简便的无有机溶剂适体固定方法开发的GFET平台检测白细胞介素-6生物标志物。

Detection of an IL-6 Biomarker Using a GFET Platform Developed with a Facile Organic Solvent-Free Aptamer Immobilization Approach.

作者信息

Khan Niazul I, Song Edward

机构信息

Department of Electrical and Computer Engineering, University of New Hampshire, Durham, NH 03824, USA.

Materials Science Program, University of New Hampshire, Durham, NH 03824, USA.

出版信息

Sensors (Basel). 2021 Feb 13;21(4):1335. doi: 10.3390/s21041335.

Abstract

Aptamer-immobilized graphene field-effect transistors (GFETs) have become a well-known detection platform in the field of biosensing with various biomarkers such as proteins, bacteria, virus, as well as chemicals. A conventional aptamer immobilization technique on graphene involves a two-step crosslinking process. In the first step, a pyrene derivative is anchored onto the surface of graphene and, in the second step, an amine-terminated aptamer is crosslinked to the pyrene backbone with EDC/NHS (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride/N-hydroxysuccinimide) chemistry. However, this process often requires the use of organic solvents such as dimethyl formamide (DMF) or dimethyl sulfoxide (DMSO) which are typically polar aprotic solvents and hence dissolves both polar and nonpolar compounds. The use of such solvents can be especially problematic in the fabrication of lab-on-a-chip or point-of-care diagnostic platforms as they can attack vulnerable materials such as polymers, passivation layers and microfluidic tubing leading to device damage and fluid leakage. To remedy such challenges, in this work, we demonstrate the use of pyrene-tagged DNA aptamers (PTDA) for performing a one-step aptamer immobilization technique to implement a GFET-based biosensor for the detection of Interleukin-6 (IL-6) protein biomarker. In this approach, the aptamer terminal is pre-tagged with a pyrene group which becomes soluble in aqueous solution. This obviates the need for using organic solvents, thereby enhancing the device integrity. In addition, an external electric field is applied during the functionalization step to increase the efficiency of aptamer immobilization and hence improved coverage and density. The results from this work could potentially open up new avenues for the use of GFET-based BioMEMS platforms by broadening the choice of materials used for device fabrication and integration.

摘要

适配体固定化的石墨烯场效应晶体管(GFET)已成为生物传感领域中一种知名的检测平台,可用于检测各种生物标志物,如蛋白质、细菌、病毒以及化学物质。传统的石墨烯适配体固定技术涉及两步交联过程。第一步,将芘衍生物锚定在石墨烯表面;第二步,使用1-乙基-3-(3-二甲基氨基丙基)碳二亚胺盐酸盐/ N-羟基琥珀酰亚胺(EDC/NHS)化学方法将胺基末端的适配体与芘主链交联。然而,这个过程通常需要使用有机溶剂(如二甲基甲酰胺(DMF)或二甲基亚砜(DMSO)),它们通常是极性非质子溶剂,因此既能溶解极性化合物,也能溶解非极性化合物。在芯片实验室或即时诊断平台的制造中,使用这类溶剂可能会特别成问题,因为它们会侵蚀诸如聚合物、钝化层和微流控管道等易损材料,导致设备损坏和液体泄漏。为了解决这些挑战,在这项工作中,我们展示了使用芘标记的DNA适配体(PTDA)来执行一步适配体固定技术,以实现用于检测白细胞介素-6(IL-6)蛋白质生物标志物的基于GFET的生物传感器。在这种方法中,适配体末端预先用芘基团标记,使其可溶于水溶液。这就无需使用有机溶剂,从而提高了设备的完整性。此外,在功能化步骤中施加外部电场,以提高适配体固定的效率,进而改善覆盖率和密度。这项工作的结果可能会通过拓宽用于设备制造和集成的材料选择,为基于GFET的生物微机电系统平台的应用开辟新途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c937/7918451/3460a9e9191e/sensors-21-01335-g0A1.jpg

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