Monitoring of the Enzymatic Reactions Course by Differential Microwave Dielectrometry Method in Real Time
Keywords:high loss liquids, complex permittivity, enzymatic reactions, electromagnetic wave propagation, differential microwave dielectrometry, enzyme trypsin, human serum albumin, immunoglobulin G
Enzymatic reactions are the basis of many biotechnological manufacturing and biomedical diagnostic procedures that require effective methods of monitoring over the reaction course. In the current paper, we present the results of the development of a new approach within the differential microwave dielectrometry method for the non-invasive monitoring of the course of enzymatic reactions based on the complex permittivity changes of these reactive mixture solutions in real time at a fixed frequency of 31.82 GHz. The dynamic studies of the dielectric parameters of selected enzymatic systems containing a protein substrate (immunoglobulin G, human serum albumin) and enzyme trypsin. The developed differential microwave dielectrometry setup has been performed to verify the proposed approach effectiveness for the enzymatic reaction monitoring in biomedical practice and the food industry. Our microwave dielectrometry results have been validated by the results of the UV-Vis spectrophotometry method for selected enzymatic systems. We propose a new approach to use the differential microwave dielectrometry method with high sensitivity (in average 0.5% and 3÷5% for the real and imaginary parts of the complex permittivity, respectively) to estimate the course of enzymatic reactions in real time.
G.D. Najafpour. Biochemical Engineering and Biotechnology (Elsevier, 2015) [ISBN: 978-0-444-63357-6].
K.S. Siddiqui, H. Ertan, A. Poljak, W.J. Bridge. Evaluating enzymatic productivity-the missing link to enzyme utility. Int. J. Mol. Sci. 23 (13), 6908 (2022).
M. Faure, M. Kechadi, B. Sotta, J. Gamby, B. Tribolleta. Contact free impedance methodology for investigating enzymatic reactions into dielectric polymer microchip. Electroanalysis 25 (5), 1151 (2013).
Y. Iwasaki, T. Horiuchi, O. Niwa. Detection of electrochemical enzymatic reactions by surface plasmon resonance measurement. Anal. Chem. 73 (7), 1595 (2001).
A. Furukawa, T. Nagata, A. Matsugami, Y. Habu, R. Sugiyama, F. Hayashi, N. Kobayashi, S. Yokoyama, H. Takaku, M. Katahira. Structure, interaction and realtime monitoring of the enzymatic reaction of wild-type APOBEC3G. The EMBO J. 28 (4), 440 (2009).
F. C. Church, H. E. Swaisgood, D.H. Porter, G.L. Cattignani. Spectrophotometric assay using o-phthaldialdehyde for determination of proteolysis in milk and isolated milk proteins. J. Dairy Sci. 66, 1219 (1983).
W.J. Ellison, K. Lamkaourchi, M.J. Moreau. Water: A dielectric reference. J. Mol. Liq. 68, 171 (1996).
A. Godio. Open ended-coaxial cable measurements of saturated sandy soils. American J. Environmental Sci. 3 (3), 175 (2007).
J. M. McKee, B.P. Johnson. Real-time chemical sensing of aqueous ethanol glucose mixtures. IEEE Transactions on Instrumentation and Measurement 49 (1), 114 (2000).
I. Dilman, M. N. Akinci, T. Yilmaz, M. ¸Cay¨oren, I. Akduman. A method to measure complex dielectric permittivity with open-ended coaxial probes. IEEE Transactions on Instrumentation and Measurement 71, (2022).
Keysight Technologies, Keysight n1501a Dielectric Probe Kit 200 MHz to 50 GHz. 2021. [Online]. Available: http://literature.cdn.keysight.com/litweb/pdf/5989-0222EN.pdf.
F. Artis, T. Chen, T. Chretiennot, J.-J. Fournie, M. Poupot, D. Dubuc, K. Grenier. Microwaving biological cells: intracellular analysis with microwave dielectric spectroscopy. IEEE Microwave Magazine 16 (4), 87 (2015).
K. Shibata. Measurement of complex permittivity for liquid materials using the open-ended cut-off waveguide reflection method. IEICE Transactions on Electronics E93-C (11), 1621 (2010).
Y. Wang, M.N. Afsar. Measurement of complex permittivity of liquids using waveguide techniques. Progress in Electromagnetics Research 42, 131 (2003).
J. Barthel, K. Bachhuber, R. Buchner, H. Hetzenauer. Dielectric spectra of some common solvents in the microwave region. Water and lower alcohols. Chem. Phys. Lett. 165 (4), 369 (1990).
Z.E. Eremenko, K.S. Kuznetsova, N.I. Sklyar, A.V. Martynov. Measuring Complex Permittivity of High-Loss Liquids in Dielectric Materials and Applications. Edited by P.G. Choudhry (Nova Science Publishers, 2019), Chap. 2 [ISBN: 978-1-53615-316-3].
Z.E. Eremenko, V.N. Skresanov, A.I. Shubnyi, N.S. Anikina, V.G. Gerzhikova, T.A. Zhilyakova. Complex Permittivity Measurement of High Loss Liquids and its Application to Wine Analysis in Electromagnetic Waves. Edited by V. Zhurbenko (Janeza Trdine, 2011) [ISBN: 978-953-307-304-0].
Z.E. Eremenko, V.A. Pashynska, K.S. Kuznetsova, O.I. Shubnyi, N.I. Sklyar, A.V. Martynov. Microwave dielectrometer application to antibiotic concentration control in water solution. Low Temperature Physics 26, (3), 30 (2021).
V.N. Skresanov, Z.E. Eremenko, E.S. Kuznetsova, Y. Wu, Y. He. Circular layered waveguide use for wideband complex permittivity measurement of lossy liquids. IEEE Transactions on Instrumentation and Measurement 63 (3), 694 (2014).
S. Schmidt, M. Schubler. All Liquid Based Calibration Scheme for Microwave Dielectrometry. In Proceedings IEEE/MTT-S International Microwave Symposium USA, 8058839 (2017).
A.A. Abduljabar, D.J. Rowe, A. Porch, D.A. Barrow. Novel microwave microfluidic sensor using a microstrip split-ring resonator. IEEE Transactions on Microwave Theory and Techniques 62 (3), 679 (2014).
A.I. Gubin, A.A. Barannik, N.T. Cherpak, I.A. Protsenko, S. Pud, A. Offenh¨ausser, S.A. Vitusevich. Whisperinggallery-mode resonator technique with microfluidic channel for permittivity measurement of liquids. IEEE Transactions on Microwave Theory and Techniques 63 (6), 2003 (2015).
J.-W.C. Alffenaar, E.M. Jongedijk, C.A.J. van Winkel, M. Sariko, S.K. Heysell, S. Mpagama, D.J. Touw. A mobile microvolume UV/visible light spectrophotometer for the measurement of levofloxacin in saliva. J. Antimicrobial Chemotherapy 76 (2), 423 (2021).
Huffman, K. Soni, J. Ferraiolo. UV-Vis based determination of protein concentration: validating and implementing slope measurements using variable pathlength technology. BioProcess International 12 (8), (2014).
How to Cite
License to Publish the Paper
The corresponding author and the co-authors (hereon referred to as the Author(s)) of the paper being submitted to the Ukrainian Journal of Physics (hereon referred to as the Paper) from one side and the Bogolyubov Institute for Theoretical Physics, National Academy of Sciences of Ukraine, represented by its Director (hereon referred to as the Publisher) from the other side have come to the following Agreement:
1. Subject of the Agreement.
The Author(s) grant(s) the Publisher the free non-exclusive right to use the Paper (of scientific, technical, or any other content) according to the terms and conditions defined by this Agreement.
2. The ways of using the Paper.
2.1. The Author(s) grant(s) the Publisher the right to use the Paper as follows.
2.1.1. To publish the Paper in the Ukrainian Journal of Physics (hereon referred to as the Journal) in original language and translated into English (the copy of the Paper approved by the Author(s) and the Publisher and accepted for publication is a constitutive part of this License Agreement).
2.1.2. To edit, adapt, and correct the Paper by approval of the Author(s).
2.1.3. To translate the Paper in the case when the Paper is written in a language different from that adopted in the Journal.
2.2. If the Author(s) has(ve) an intent to use the Paper in any other way, e.g., to publish the translated version of the Paper (except for the case defined by Section 2.1.3 of this Agreement), to post the full Paper or any its part on the web, to publish the Paper in any other editions, to include the Paper or any its part in other collections, anthologies, encyclopaedias, etc., the Author(s) should get a written permission from the Publisher.
3. License territory.
The Author(s) grant(s) the Publisher the right to use the Paper as regulated by sections 2.1.1–2.1.3 of this Agreement on the territory of Ukraine and to distribute the Paper as indispensable part of the Journal on the territory of Ukraine and other countries by means of subscription, sales, and free transfer to a third party.
4.1. This Agreement is valid starting from the date of signature and acts for the entire period of the existence of the Journal.
5.1. The Author(s) warrant(s) the Publisher that:
– he/she is the true author (co-author) of the Paper;
– copyright on the Paper was not transferred to any other party;
– the Paper has never been published before and will not be published in any other media before it is published by the Publisher (see also section 2.2);
– the Author(s) do(es) not violate any intellectual property right of other parties. If the Paper includes some materials of other parties, except for citations whose length is regulated by the scientific, informational, or critical character of the Paper, the use of such materials is in compliance with the regulations of the international law and the law of Ukraine.
6. Requisites and signatures of the Parties.
Publisher: Bogolyubov Institute for Theoretical Physics, National Academy of Sciences of Ukraine.
Address: Ukraine, Kyiv, Metrolohichna Str. 14-b.
Author: Electronic signature on behalf and with endorsement of all co-authors.