Digital microfluidic immobilized cytochrome P450 reactors with integrated inkjet-printed microheaters for droplet-based drug metabolism research

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http://hdl.handle.net/10138/245422

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Sathyanarayanan , G , Haapala , M , Kiiski , I & Sikanen , T 2018 , ' Digital microfluidic immobilized cytochrome P450 reactors with integrated inkjet-printed microheaters for droplet-based drug metabolism research ' , Analytical and Bioanalytical Chemistry , vol. 410 , no. 25 , pp. 6677-6687 . https://doi.org/10.1007/s00216-018-1280-7

Title: Digital microfluidic immobilized cytochrome P450 reactors with integrated inkjet-printed microheaters for droplet-based drug metabolism research
Author: Sathyanarayanan, Gowtham; Haapala, Markus; Kiiski, Iiro; Sikanen, Tiina
Contributor: University of Helsinki, Preclinical Drug Formulation and Analysis group
University of Helsinki, Division of Pharmaceutical Chemistry and Technology
University of Helsinki, Faculty of Pharmacy
University of Helsinki, Faculty of Pharmacy
Date: 2018-10
Language: eng
Number of pages: 11
Belongs to series: Analytical and Bioanalytical Chemistry
ISSN: 1618-2650
URI: http://hdl.handle.net/10138/245422
Abstract: We report the development and characterization of digital microfluidic (DMF) immobilized enzyme reactors (IMERs) for studying cytochrome P450 (CYP)-mediated drug metabolism on droplet scale. The on-chip IMERs consist of porous polymer (thiol-ene) monolith plugs prepared in situ by photopolymerization and functionalized with recombinant CYP1A1 isoforms (an important detoxification route for many drugs and other xenobiotics). The DMF devices also incorporate inexpensive, inkjet-printed microheaters for on-demand regio-specific heating of the IMERs to physiological temperature, which is crucial for maintaining the activity of the temperature-sensitive CYP reaction. For on-chip monitoring of the CYP activity, the DMF devices were combined with a commercial well-plate reader, and a custom fluorescence quantification method was developed for detection of the chosen CYP1A1 model activity (ethoxyresorufin-O-deethylation). The reproducibility of the developed assay was examined with the help of ten parallel CYP-IMERs. All CYP-IMERs provided statistically significant difference (in fluorescence response) compared to any of the negative controls (including room-temperature reactions). The average (n = 10) turnover rate was 20.3 +/- 9.0 fmol resorufin per minute. Via parallelization, the concept of the droplet-based CYP-IMER developed in this study provides a viable approach to rapid and low-cost prediction of the metabolic clearance of new chemical entities in vitro.
Subject: 317 Pharmacy
Digital microfluidics
Drug metabolism
Cytochrome P450
Microheater
Microreactor
Enzyme immobilization
TOOL
CHIP
DEVICES
PLATFORM
STABILITY
MECHANISMS
BIOLOGY
HIGH-THROUGHPUT
1182 Biochemistry, cell and molecular biology
116 Chemical sciences
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