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        Electrowetting-Based Digital Microfluidics Platform for Automated Enzyme-linked Immunosorbent Assay

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        jove_protocol_60489_electrowetting_based_digital_microfluidics_platform_for_automated.pdf (PDF, 1Mb)
        Author
        Dimov, Nikolay
        McDonnell, M.B.
        Munro, Ian
        McCluskey, Daniel
        Johnston, Ian
        Tan, Christabel
        Coudron, Loic
        Attention
        2299/22356
        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 H 2O 2/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.
        Publication date
        2020-02-23
        Published in
        Journal of Visualized Experiments
        Published version
        https://doi.org/10.3791/60489
        Other links
        http://hdl.handle.net/2299/22356
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