Mendel University in Brno - list of publications


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NUCHTAVORN, N. -- LEANPOLCHAREANCHAI, J. -- SUNTORNSUK, L. -- MACKA, M. Paper-based sol-gel thin films immobilized cytochrome P450 for enzyme activity measurement. Analytica Chimica Acta. 2020. v. 1098, no. 15 February, p. 86--93. ISSN 0003-2670. URL: https://doi.org/10.1016/j.aca.2019.11.031

Original name: Paper-based sol-gel thin films immobilized cytochrome P450 for enzyme activity measurement
Czech name:
Written by (author): Nantana Nuchtavorn
Jiraporn Leanpolchareanchai
Leena Suntornsuk
doc. RNDr. Miroslav Macka, Ph.D.
Department: Department of Chemistry and Biochemistry
Kind of publication: article in a professional periodical
Periodical: Analytica Chimica Acta
Nature of article: paper
Volume no. (year): 1098
Periodical number within the volume: 15 February
Year of publication: 2020
Starting page: 86
Up to page: 93
Number of pages: 8
Sub-specification: článek je obsažen v databázi Web of Science
UT code by Web of Science:
EID code by Scopus:
Form of publication: printed version
Original language: English
Description in original language: Cytochrome P450 (CYP450), and in particular CYP3A4, is the most abundantly expressed CYP450 isozyme implicated in many drug-drug and medicinal plant-drug interactions. Therefore, incorporation of CYP3A4 enzyme screening at an early stage of drug discovery is preferable in order to avoid enzymatic interactions. Here we present for the first time a paper-based CYP3A4 immobilized sol-gel-derived a platform using resorufin benzyl ether as a fluorogenic enzyme substrate used to investigate enzyme activity. The fluorescence intensity of the product can be simply quantified by using a handheld digital microscope and an image analysis software. The limit of quantitation was 0.35 μM with good precision (RSDs < 4.1%). Furthermore, the assay of CYP3A4 activity on the developed paper-based device provided comparable results with those obtained from conventional well-plates (p > 0.05), while offering simplicity and lower cost. Kinetic parameters of the immobilized CYP3A4 in sol-gel coated paper were calculated from the Lineweaver-Burk plot, including Michaelis constant (Km) and maximum velocity (Vmax), which were 2.71 +- 0.35 μM and 0.43 +- 0.05 μM/min, respectively. Moreover, a functional test of these devices was conducted by assessments of known CYP3A4 inhibitors (i.e. ketoconazole, itraconazole) and inducers (i.e. phenytoin, carbamazepine). To further demonstrate the broad range of uses, the devices were utilized to assay plant extracts i.e. Areca catechu seeds, Camellia sinensis leaves, Eclipta prostrata aerial part, providing results in good agreement with previous studies. Furthermore, the sol-gel immobilized enzyme stored at 4 oC can increase storage stability, offering the activity of 86.3 +- 0.4% after 3-weeks storage, equivalent to the activity of the free enzyme solution after 1-week storage. The developed paper-based devices offer versatility, portability and low-cost.
Description in English:
Description in Czech:
Year of submission: 2020
Year of transmission:
RIV identification number:
URL: https://doi.org/10.1016/j.aca.2019.11.031
 
Entry made by: Markéta Hejčová, DiS.
Last change: 01/20/2020 10:43 (Markéta Hejčová, DiS.)

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Source specification:

Analytica Chimica Acta. ISSN 0003-2670.

Original name: Analytica Chimica Acta
English name:
Czech name:
Written by (author):
Kind of publication: magazine
ISSN: 0003-2670
Country of publisher: Kingdom of the Netherlands
Place of publishing:
Publisher: Elsevier Science BV
URL:
Reviewed magazine: no
Original language:
Description in original language:
Description in English:
Description in Czech:
 
Entry made by: Lenka Baláková
Last change: 03/14/2012 20:37 (Ing. Aleš Kutín)

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