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Co-encapsulated urease and glucose oxidase in polyelectrolyte microcapsules for simultaneous glucose and urea determination.

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Τίτλος: Co-encapsulated urease and glucose oxidase in polyelectrolyte microcapsules for simultaneous glucose and urea determination.
Συγγραφείς: Kim AL; Institute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya St., 3, Puschino, Moscow Region, 142290, Russia. Electronic address: kimerzent@gmail.com., Musin EV; Institute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya St., 3, Puschino, Moscow Region, 142290, Russia. Electronic address: eglork@gmail.com., Tikhonenko SA; Institute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya St., 3, Puschino, Moscow Region, 142290, Russia. Electronic address: tikhonenkosa@gmail.com.
Πηγή: Analytica chimica acta [Anal Chim Acta] 2026 Oct 15; Vol. 1419, pp. 345902. Date of Electronic Publication: 2026 Jun 30.
Τύπος έκδοσης: Journal Article; Research Support, Non-U.S. Gov't
Γλώσσα: English
Στοιχεία περιοδικού: Publisher: Elsevier Country of Publication: Netherlands NLM ID: 0370534 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-4324 (Electronic) Linking ISSN: 00032670 NLM ISO Abbreviation: Anal Chim Acta Subsets: MEDLINE
Imprint Name(s): Publication: Amsterdam : Elsevier
Original Publication: Amsterdam.
Ιατρικοί όροι (MeSH): Glucose Oxidase*/chemistry , Glucose Oxidase*/metabolism , Urease*/chemistry , Urease*/metabolism , Urea*/analysis , Glucose*/analysis , Polyelectrolytes*/chemistry, Capsules/chemistry ; Enzymes, Immobilized/chemistry ; Enzymes, Immobilized/metabolism ; Humans
Περίληψη: Background: Clinical monitoring of metabolic disorders requires stable tools for simultaneous analyte detection. However, conventional free-enzyme assays suffer from rapid degradation, poor operational stability, and single-use limitations. Herein, we report a novel bi-enzymatic diagnostic system based on the rational co-encapsulation of glucose oxidase (GOx) and urease within polyelectrolyte microcapsules (PMCs) for the simultaneous determination of glucose and urea.
Results: Fabricated via layer-by-layer assembly, the PMCs achieved high enzyme loading (73 ± 2% for GOx and 93 ± 5% for urease). Co-encapsulation significantly reduced the GOx Michaelis constant from 10.1 to 2.5 mM. The system exhibited linear ranges of 0.5-3.0 mM (glucose) and 1-40 mM (urea), covering clinically relevant concentrations. It maintained performance for over 61 measurement cycles and a 27-day shelf-life at room temperature. Direct analysis of human saliva revealed that the PMCs effectively shield enzymes from matrix interferences, yielding significantly lower data dispersion and higher reproducibility than standard Clark-electrode and Berthelot reference methods.
Significance: By combining multi-cycle reusability, room-temperature stability, and enhanced precision in raw biological fluids, this co-encapsulation strategy overcomes the fundamental trade-offs of traditional immobilization. The developed PMCs provide a robust, cost-effective, and comprehensive platform for point-of-care metabolic diagnostics and non-invasive monitoring, establishing a generalizable framework for future multi-enzymatic biosensing applications.
(Copyright © 2026 Elsevier B.V. All rights reserved.)
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Contributed Indexing: Keywords: Bienzymatic; Co-encapsulation; Diagnostic system; Glucose oxidase; Multiplex detection; Polyelectrolyte microcapsules; Urease
Substance Nomenclature: EC 1.1.3.4 (Glucose Oxidase)
EC 3.5.1.5 (Urease)
8W8T17847W (Urea)
0 (Capsules)
IY9XDZ35W2 (Glucose)
0 (Enzymes, Immobilized)
0 (Polyelectrolytes)
Entry Date(s): Date Created: 20260819 Date Completed: 20260819 Latest Revision: 20260827
Update Code: 20260827
DOI: 10.1016/j.aca.2026.345902
PMID: 42618110
Βάση Δεδομένων: MEDLINE
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  Data: Co-encapsulated urease and glucose oxidase in polyelectrolyte microcapsules for simultaneous glucose and urea determination.
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  Data: <searchLink fieldCode="AU" term="%22Kim+AL%22">Kim AL</searchLink>; Institute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya St., 3, Puschino, Moscow Region, 142290, Russia. Electronic address: kimerzent@gmail.com.<br /><searchLink fieldCode="AU" term="%22Musin+EV%22">Musin EV</searchLink>; Institute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya St., 3, Puschino, Moscow Region, 142290, Russia. Electronic address: eglork@gmail.com.<br /><searchLink fieldCode="AU" term="%22Tikhonenko+SA%22">Tikhonenko SA</searchLink>; Institute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya St., 3, Puschino, Moscow Region, 142290, Russia. Electronic address: tikhonenkosa@gmail.com.
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  Data: <searchLink fieldCode="JN" term="%220370534%22">Analytica chimica acta</searchLink> [Anal Chim Acta] 2026 Oct 15; Vol. 1419, pp. 345902. <i>Date of Electronic Publication: </i>2026 Jun 30.
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  Data: Journal Article; Research Support, Non-U.S. Gov't
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  Data: English
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  Data: <i>Publisher: </i><searchLink fieldCode="PB" term="%22Elsevier%22">Elsevier </searchLink><i>Country of Publication: </i>Netherlands <i>NLM ID: </i>0370534 <i>Publication Model: </i>Print-Electronic <i>Cited Medium: </i>Internet <i>ISSN: </i>1873-4324 (Electronic) <i>Linking ISSN: </i><searchLink fieldCode="IS" term="%2200032670%22">00032670 </searchLink><i>NLM ISO Abbreviation: </i>Anal Chim Acta <i>Subsets: </i>MEDLINE
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  Data: <i>Publication</i>: Amsterdam : Elsevier<br /><i>Original Publication</i>: Amsterdam.
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  Data: <searchLink fieldCode="MM" term="%22Glucose+Oxidase%22">Glucose Oxidase*</searchLink>/<searchLink fieldCode="MM" term="%22Glucose+Oxidase+chemistry%22">chemistry</searchLink> <br /><searchLink fieldCode="MM" term="%22Glucose+Oxidase%22">Glucose Oxidase*</searchLink>/<searchLink fieldCode="MM" term="%22Glucose+Oxidase+metabolism%22">metabolism</searchLink> <br /><searchLink fieldCode="MM" term="%22Urease%22">Urease*</searchLink>/<searchLink fieldCode="MM" term="%22Urease+chemistry%22">chemistry</searchLink> <br /><searchLink fieldCode="MM" term="%22Urease%22">Urease*</searchLink>/<searchLink fieldCode="MM" term="%22Urease+metabolism%22">metabolism</searchLink> <br /><searchLink fieldCode="MM" term="%22Urea%22">Urea*</searchLink>/<searchLink fieldCode="MM" term="%22Urea+analysis%22">analysis</searchLink> <br /><searchLink fieldCode="MM" term="%22Glucose%22">Glucose*</searchLink>/<searchLink fieldCode="MM" term="%22Glucose+analysis%22">analysis</searchLink> <br /><searchLink fieldCode="MM" term="%22Polyelectrolytes%22">Polyelectrolytes*</searchLink>/<searchLink fieldCode="MM" term="%22Polyelectrolytes+chemistry%22">chemistry</searchLink><br /><searchLink fieldCode="MH" term="%22Capsules%22">Capsules</searchLink>/<searchLink fieldCode="MH" term="%22Capsules+chemistry%22">chemistry</searchLink> ; <searchLink fieldCode="MH" term="%22Enzymes%2C+Immobilized%22">Enzymes, Immobilized</searchLink>/<searchLink fieldCode="MH" term="%22Enzymes%2C+Immobilized+chemistry%22">chemistry</searchLink> ; <searchLink fieldCode="MH" term="%22Enzymes%2C+Immobilized%22">Enzymes, Immobilized</searchLink>/<searchLink fieldCode="MH" term="%22Enzymes%2C+Immobilized+metabolism%22">metabolism</searchLink> ; <searchLink fieldCode="MH" term="%22Humans%22">Humans</searchLink>
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  Data: Background: Clinical monitoring of metabolic disorders requires stable tools for simultaneous analyte detection. However, conventional free-enzyme assays suffer from rapid degradation, poor operational stability, and single-use limitations. Herein, we report a novel bi-enzymatic diagnostic system based on the rational co-encapsulation of glucose oxidase (GOx) and urease within polyelectrolyte microcapsules (PMCs) for the simultaneous determination of glucose and urea.<br />Results: Fabricated via layer-by-layer assembly, the PMCs achieved high enzyme loading (73 ± 2% for GOx and 93 ± 5% for urease). Co-encapsulation significantly reduced the GOx Michaelis constant from 10.1 to 2.5 mM. The system exhibited linear ranges of 0.5-3.0 mM (glucose) and 1-40 mM (urea), covering clinically relevant concentrations. It maintained performance for over 61 measurement cycles and a 27-day shelf-life at room temperature. Direct analysis of human saliva revealed that the PMCs effectively shield enzymes from matrix interferences, yielding significantly lower data dispersion and higher reproducibility than standard Clark-electrode and Berthelot reference methods.<br />Significance: By combining multi-cycle reusability, room-temperature stability, and enhanced precision in raw biological fluids, this co-encapsulation strategy overcomes the fundamental trade-offs of traditional immobilization. The developed PMCs provide a robust, cost-effective, and comprehensive platform for point-of-care metabolic diagnostics and non-invasive monitoring, establishing a generalizable framework for future multi-enzymatic biosensing applications.<br /> (Copyright © 2026 Elsevier B.V. All rights reserved.)
– Name: Abstract
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  Data: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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  Data: <i>Keywords: </i>Bienzymatic; Co-encapsulation; Diagnostic system; Glucose oxidase; Multiplex detection; Polyelectrolyte microcapsules; Urease
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  Data: EC 1.1.3.4 (Glucose Oxidase)<br />EC 3.5.1.5 (Urease)<br />8W8T17847W (Urea)<br />0 (Capsules)<br />IY9XDZ35W2 (Glucose)<br />0 (Enzymes, Immobilized)<br />0 (Polyelectrolytes)
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  Data: 10.1016/j.aca.2026.345902
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        Value: 10.1016/j.aca.2026.345902
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        Text: English
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      – SubjectFull: Enzymes, Immobilized chemistry
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      – SubjectFull: Urea analysis
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              Text: 2026 Oct 15
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