Single-step Trypsin Inhibitor Assay on a Microchannel Array Device Immobilizing Enzymes and Fluorescent Substrates by Inkjet Printing

• KAWAI Yuko

  Department of Applied Chemistry, Graduate School of Engineering, Osaka Prefecture University

• IDEGAMI Kotaro

  Sysmex Corporation

• SUEYOSHI Kenji

  Department of Applied Chemistry, Graduate School of Engineering, Osaka Prefecture University Japan Science and Technology Agency (JST), Precursory Research for Embryonic Science and Technology (PRESTO)

• ENDO Tatsuro

  Department of Applied Chemistry, Graduate School of Engineering, Osaka Prefecture University

• HISAMOTO Hideaki

  Department of Applied Chemistry, Graduate School of Engineering, Osaka Prefecture University

Abstract

In this paper, we report a single-step trypsin inhibitor assay on a microchannel array device immobilizing enzymes and substrates by inkjet printing. The microdevice is composed of a poly(dimethylsiloxane) (PDMS) microchannel array that immobilizes trypsin and fluorescent substrates as reactive reagents at the two bottom corners of a microchannel. Inkjet printers allow simple, accurate, and position-selective immobilization of reagents as nanoliter spots. Therefore, plural reactive reagents, such as enzymes and substrates, can be separately immobilized at different positions in the same microchannel without mixing, and thus allowing for single-step operation by simply introducing a sample solution through capillary action. Furthermore, reproducible fabrication and mass production of the device could be expected. In this study, the efficiency of an acidic solution as a spotting agent for protease immobilization to prevent decrease in the fluorescence intensity was confirmed. Additionally, single-step trypsin inhibitor screening was performed using three inhibitors. Finally, we investigated the storage stability of the device and confirmed that it remained stable for at least 10 days.

Journal

• Analytical Sciences

  Analytical Sciences 37 (10), 1473-1476, 2021-10-10

  The Japan Society for Analytical Chemistry

Keywords

• Enzyme inhibitor assay
• inkjet printing
• microchannel
• poly(dimethylsiloxane)
• single-step

Details

• CRID: 1390008212404945408
• NII Article ID: 130008102510
• NII Book ID: AA10500785
• DOI: 10.2116/analsci.21n011
• ISSN: 13482246
• ISSN: 09106340
• NDL BIB ID: 031754512
• PubMed: 33952864
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I031754512
• Web Site: https://link.springer.com/content/pdf/10.2116/analsci.21N011.pdf
• Web Site: https://link.springer.com/article/10.2116/analsci.21N011/fulltext.html
• Web Site: https://www.jstage.jst.go.jp/article/analsci/37/10/37_21N011/_pdf
• Web Site: https://search.jamas.or.jp/link/ui/2022251354
• Text Lang: en
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed