Biofabrication Using Pyrrole Electropolymerization for the Immobilization of Glucose Oxidase and Lactate Oxidase on Implanted Microfabricated Biotransducers
Abstract
:1. Introduction
2. Experimental Section
2.1. Chemicals and Reagents
2.2. Enzyme Monomerization and Sulfonization
2.3. Electropolymerization and Electrochemical Characterization
2.4. Transducer Cleaning and Seeding with Polypyrrole
2.5. Conferring Biospecificity to Polypyrrole Seeded Transducers
2.6. In Vitro Biosensor Calibration Prior to in Vivo Implantation
2.7. Implantation of Biotransducers into the Trapezius Muscles of Sprague-Dawley Rats
2.8. In Vivo Biosensor Sensitivity and Response Time to Bolus Infusion of Analytes
3. Results and Discussion
3.1. Effects of a Polypyrrole Seeding Layer on Analytical Performance of Biotransducers
3.2. Enzyme Monomerization and Sulfonization
3.3. Electropolymerization Kinetics of Covalently Conjugated Systems
3.4. Characterization of Enzyme Conjugation using Ferrocene Monocarboxylic Acid
3.5. Bioanalytical Performance of Monomerized and Sulfonated Enzyme Biosensors
[E] = 1.0 mg/mL | PPy-GOx | PPy(SBA)-GOx | PPy(SBA-con-GOx) | P(Py-co-PyBA)-GOx | P(Py-co-PyBA-con-Gox) (n = 2) |
---|---|---|---|---|---|
(n = 4) | (n = 4) | (n = 2) | (n = 2) | ||
Polymerizaton time(R.L. 100 mC/cm2) (min) | 68 ± 22 | 23 ± 19 | 15 ± 5 | 204 ± 37 | 230 ± 132 |
Sensitivity (µA·cm−2·mM−1) | 0.87 ± 0.04 | 1.8 ± 1.2 | 1.9 ± 0.2 | 4.8 ± 0.9 | 0.99 ± 0.28 |
Response time (s) | 41 ± 21 | 17 ± 9 | 18 ± 5 | 17 ± 5 | 14 ± 5 |
Detection limit (mM) | 0.5 ± 0.3 | 0.01 ± 0.01 | 2.2 × 10−3± 1.8 × 10−3 | 0.05 ± 0.03 | 0.02 ± 0.01 |
KMapp (mM) | 14.1 ± 3.1 | 8.2 ± 2.6 | 11.1 ± 5.6 | 7.2 ± 2.6 | 19.7 ± 3.9 |
Imax (µA·cm−2) | 23.3 ± 3.1 | 30.6 ± 18.8 | 39.2 ± 11.7 | 77 ± 25 | 28.9 ± 4.0 |
Linear range (mM) | 17.4 ± 7.0 | 9.4 ± 2.6 | 10.2 ± 3.2 | 9.5 ± 2.8 | 25.6 ± 5.6 |
3.6. Effect of Extent of Monomerizaton or Sulfonation
3.7. Performance of Freshly Prepared and Resected Biotransducers
MDEA 5037-Pt|PPy(PSSA)|p(Py-co-PyBA)-LOx | MDEA 5037-Pt|PPy(PSSA)|p(Py-co-PyBA)-GOx | |||
---|---|---|---|---|
[E] = 1.0 mg/mL | [E] = 1.0 mg/mL | |||
Freshly prepared biotransducers | Resected biotransducers | Freshly prepared biotransducers | Resected biotransducers | |
(n = 3) | (n = 3) | (n = 4) | (n = 4) | |
Sensitivity (µA·cm−2·mM−1) | 0.36 ± 0.15 | - | 0.68 ± 0.40 | 0.22 ± 0.17 |
Response time (s) | 86 ± 26 | - | 41 ± 18 | 244 ± 193 |
Detection limit (mM) | 7.9 ± 5.6 | - | 0.05 ± 0.03 | 0.27 ± 0.27 |
KMapp (mM) | −9.1 ± 15.8 | - | 7.3 ± 3.4 | 74 ± 130 |
Imax (µA·cm−2) | −1.9 ± 5.8 | - | 11.7 ± 6.5 | 8.8 ± 9.3 |
Linear range (mM) | 6.6 ± 0.9 | - | 7.2 ± 5.9 | 14.1 ± 11.6 |
3.8. In Vivo Response of MDEA 5037 Biotransducers to Exogenously Changing Analytes
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Kotanen, C.N.; Karunwi, O.; Guiseppi-Elie, A. Biofabrication Using Pyrrole Electropolymerization for the Immobilization of Glucose Oxidase and Lactate Oxidase on Implanted Microfabricated Biotransducers. Bioengineering 2014, 1, 85-110. https://doi.org/10.3390/bioengineering1010085
Kotanen CN, Karunwi O, Guiseppi-Elie A. Biofabrication Using Pyrrole Electropolymerization for the Immobilization of Glucose Oxidase and Lactate Oxidase on Implanted Microfabricated Biotransducers. Bioengineering. 2014; 1(1):85-110. https://doi.org/10.3390/bioengineering1010085
Chicago/Turabian StyleKotanen, Christian N., Olukayode Karunwi, and Anthony Guiseppi-Elie. 2014. "Biofabrication Using Pyrrole Electropolymerization for the Immobilization of Glucose Oxidase and Lactate Oxidase on Implanted Microfabricated Biotransducers" Bioengineering 1, no. 1: 85-110. https://doi.org/10.3390/bioengineering1010085