Single-drop analysis of various proteases in a cancer cell lysate using a capillary-assembled microchip |
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Authors: | Terence G Henares Fumio Mizutani Ryuichi Sekizawa Hideaki Hisamoto |
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Institution: | (1) Graduate School of Material Science, University of Hyogo, 3–2–1 Kouto, Kamigori-cho, Ako-gun Hyogo, 678–1297, Japan;(2) Metaboscreen Co. Ltd., 34–1–412, Terakubo Naka-ku, Yokohama 231–0855, Japan;(3) Graduate School of Engineering, Osaka Prefecture University, 1–1 Gakuen-cho, Naka-ku, Sakai City, Osaka 599–8531, Japan |
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Abstract: | Single-drop analysis of two different real sample solutions (2 μL) while simultaneously monitoring the activity of two sets
of ten different proteases on a single microfluidic device is presented. The device, called a capillary-assembled microchip
(CAs-CHIP), is fabricated by embedding square glass sensing capillaries (reagent-release capillaries, RRC) in the polydimethylsiloxane
(PDMS) lattice microchannel, and used for that purpose. First, the performance reliability was evaluated by measuring the
fluorescence response of twenty caspase-3-sensing capillaries on a single CAs-CHIP, and a relative standard deviation of 1.5–8.2
(% RSD, n = 5 or 10) was obtained. This suggests that precise multiplexed protease-activity sensing is possible by using a single CAs-CHIP
with multiple RRCs embedded. Then, using a single CAs-CHIP, real sample analysis of the activity of ten different caspases/proteases
in cervical cancer (HeLa) cell lysate treated and untreated with the cell-death-inducer drug, doxorubicin, was simultaneously
carried out, and a significant difference in enzyme activity between these two samples was observed. These results suggested
the usefulness of the CAs-CHIP in the field of drug discovery.
Figure Single drop analysis of two real sample solutions including various different proteases using a single microfluidic device
was achieved |
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Keywords: | Capillary-assembled microchip HeLa cell lysate Multiplexed protease-activity sensing Reagent-release capillary |
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