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Biosensors
Volume 15
Issue 2
10.3390/bios15020077
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Enzyme Biosensor Based on 3D-Printed Flow-Through Reactor Modified with Thiacalixarene-Functionalized Oligo (Lactic Acids)

by
Dmitry Stoikov
1,
Dominika Kappo
1,
Alexey Ivanov
1,*,
Vladimir Gorbachuk
1,
Olga Mostovaya
1,
Pavel Padnya
1,
Ivan Stoikov
1 and
Gennady Evtugyn
1,2,†
1
Alexander Butlerov Institute of Chemistry, Kazan Federal University, 18 Kremlevskaya Street, Kazan 420008, Russia
2
Analytical Chemistry Department, Chemical Technology Institute, Ural Federal University, 19 Mira Street, Ekaterinburg 620002, Russia
*
Author to whom correspondence should be addressed.
†
Deceased author.
Biosensors 2025, 15(2), 77; https://doi.org/10.3390/bios15020077
Submission received: 13 December 2024 / Revised: 26 January 2025 / Accepted: 28 January 2025 / Published: 29 January 2025
(This article belongs to the Special Issue Feature Paper in Biosensor and Bioelectronic Devices 2024)
Versions Notes

Abstract

Electrochemical enzyme biosensors are extensively utilized in clinical analysis and environmental monitoring, yet achieving effective enzyme immobilization while maintaining high activity remains a challenge. In this work, we developed a flow-through enzyme biosensor system using a 3D-printed flow-through electrochemical cell fabricated from commercially available poly (lactic acid). After modification with thiacalixarene-functionalized oligo (lactic acids) (OLAs), the material enabled efficient immobilization of uricase on the inner surface of a replaceable reactor of the cell. Swelling and hydrolytic stability of OLAs in cone, partial cone, and 1,3-alternate conformations were studied, with 1,3-alernate conformation demonstrating superior stability and enzyme immobilization performance. The use of OLAs enhanced immobilization efficiency by over 30% and protected the reactor from swelling, hydrolytic degradation, and enzyme loss. The biosensor was validated for amperometric uric acid determination, with a screen-printed carbon electrode modified with carbon black and Prussian Blue. This modification reduced the cathodic potential for uric acid detection to –0.05 V. The biosensor exhibited a linear detection range of 10 nM to 30 μM with a detection limit of 7 nM, and it performed effectively in artificial urine and synthetic blood plasma. The novel cell design, featuring easy assembly and low-cost replaceable parts, makes this biosensor a promising candidate for routine clinical analysis and other practical applications.
Keywords: flow-through analysis; chronoamperometry; electrochemical biosensor; replaceable reactor; uricase; thiacalixarene-functionalized oligo (lactic acids) flow-through analysis; chronoamperometry; electrochemical biosensor; replaceable reactor; uricase; thiacalixarene-functionalized oligo (lactic acids)

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MDPI and ACS Style

Stoikov, D.; Kappo, D.; Ivanov, A.; Gorbachuk, V.; Mostovaya, O.; Padnya, P.; Stoikov, I.; Evtugyn, G. Enzyme Biosensor Based on 3D-Printed Flow-Through Reactor Modified with Thiacalixarene-Functionalized Oligo (Lactic Acids). Biosensors 2025, 15, 77. https://doi.org/10.3390/bios15020077

AMA Style

Stoikov D, Kappo D, Ivanov A, Gorbachuk V, Mostovaya O, Padnya P, Stoikov I, Evtugyn G. Enzyme Biosensor Based on 3D-Printed Flow-Through Reactor Modified with Thiacalixarene-Functionalized Oligo (Lactic Acids). Biosensors. 2025; 15(2):77. https://doi.org/10.3390/bios15020077

Chicago/Turabian Style

Stoikov, Dmitry, Dominika Kappo, Alexey Ivanov, Vladimir Gorbachuk, Olga Mostovaya, Pavel Padnya, Ivan Stoikov, and Gennady Evtugyn. 2025. "Enzyme Biosensor Based on 3D-Printed Flow-Through Reactor Modified with Thiacalixarene-Functionalized Oligo (Lactic Acids)" Biosensors 15, no. 2: 77. https://doi.org/10.3390/bios15020077

APA Style

Stoikov, D., Kappo, D., Ivanov, A., Gorbachuk, V., Mostovaya, O., Padnya, P., Stoikov, I., & Evtugyn, G. (2025). Enzyme Biosensor Based on 3D-Printed Flow-Through Reactor Modified with Thiacalixarene-Functionalized Oligo (Lactic Acids). Biosensors, 15(2), 77. https://doi.org/10.3390/bios15020077

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