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用于自动化酶联免疫吸附测定的基于电润湿的数字微流控平台。

Electrowetting-based Digital Microfluidics Platform for Automated Enzyme-linked Immunosorbent Assay.

作者信息

Dimov Nikolay, McDonnell Martin B, Munro Ian, McCluskey Daniel K, Johnston Ian D, Tan Christabel K L, Coudron Loïc

机构信息

School of Engineering and Computer Science, University of Hertfordshire.

School of Engineering and Computer Science, University of Hertfordshire;

出版信息

J Vis Exp. 2020 Feb 23(156). doi: 10.3791/60489.

Abstract

Electrowetting is the effect by which the contact angle of a droplet exposed to a surface charge is modified. Electrowetting-on-dielectric (EWOD) exploits the dielectric properties of thin insulator films to enhance the charge density and hence boost the electrowetting effect. The presence of charges results in an electrically induced spreading of the droplet which permits purposeful manipulation across a hydrophobic surface. Here, we demonstrate EWOD-based protocol for sample processing and detection of four categories of antigens, using an automated surface actuation platform, via two variations of an Enzyme-Linked Immunosorbent Assay (ELISA) methods. The ELISA is performed on magnetic beads with immobilized primary antibodies which can be selected to target a specific antigen. An antibody conjugated to HRP binds to the antigen and is mixed with H2O2/Luminol for quantification of the captured pathogens. Assay completion times of between 6 and 10 min were achieved, whilst minuscule volumes of reagents were utilized.

摘要

电润湿是指暴露于表面电荷的液滴接触角发生改变的效应。介电上电润湿(EWOD)利用薄绝缘膜的介电特性来提高电荷密度,从而增强电润湿效应。电荷的存在导致液滴发生电诱导铺展,从而可以在疏水表面上进行有目的的操控。在此,我们展示了一种基于EWOD的方案,该方案使用自动化表面驱动平台,通过酶联免疫吸附测定(ELISA)方法的两种变体,对四类抗原进行样品处理和检测。ELISA在固定有一抗的磁珠上进行,一抗可被选择用于靶向特定抗原。与辣根过氧化物酶(HRP)偶联的抗体与抗原结合,并与过氧化氢/鲁米诺混合,用于定量捕获的病原体。检测完成时间在6至10分钟之间,同时试剂用量极少。

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