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

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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 organization: Preclinical Drug Formulation and Analysis group
Faculty of Pharmacy
Division of Pharmaceutical Chemistry and Technology
Drug Research Program
Tiina Sikanen / Chemical Microsystems Lab
Date: 2018-10
Language: eng
Number of pages: 11
Belongs to series: Analytical and Bioanalytical Chemistry
ISSN: 1618-2650
DOI: https://doi.org/10.1007/s00216-018-1280-7
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
Peer reviewed: Yes
Rights: cc_by
Usage restriction: openAccess
Self-archived version: publishedVersion


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