Single-drop analysis of various proteases in a cancer cell lysate using a capillary-assembled microchip

Analytical and Bioanalytical Chemistry
Terence G HenaresHideaki Hisamoto

Abstract

Single-drop analysis of two different real sample solutions (2 microL) 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.

References

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May 26, 2005·Lab on a Chip·Cristina Muñoz-PinedoAlbert van den Berg
Sep 10, 2005·Electrophoresis·Jianhua QinBingcheng Lin
Apr 28, 2006·Analytical and Bioanalytical Chemistry·Hideaki HisamotoShigeru Terabe
Aug 29, 2007·Analytica Chimica Acta·Hideaki HisamotoShigeru Terabe
Jan 12, 2008·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Terence G HenaresHideaki Hisamoto

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Citations

Jan 11, 2012·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Erina TsutsumiHideaki Hisamoto
Sep 21, 2010·Analytical Biochemistry·Urja S LathiaMark Nitz
Nov 17, 2009·Analytical Biochemistry·Urja S LathiaMark Nitz
Jan 15, 2014·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Terence G HenaresHideaki Hisamoto
Nov 13, 2015·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Kenji SueyoshiHideaki Hisamoto

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