An amperometric urea biosensor based on covalently immobilized urease on an electrode made of hyperbranched polyester functionalized gold nanoparticles

Ashutosh Tiwari; Santosh Aryal; Srikanth Pilla; Shaoqin Gong

doi:10.1016/j.talanta.2009.02.038

An amperometric urea biosensor based on covalently immobilized urease on an electrode made of hyperbranched polyester functionalized gold nanoparticles

Ashutosh Tiwari, Santosh Aryal, Srikanth Pilla, Shaoqin Gong

Houston Methodist

Research output: Contribution to journal › Article › peer-review

112 Scopus citations

Abstract

An amperometric biosensor was fabricated for the quantitative determination of urea in aqueous medium using hematein, a pH-sensitive natural dye. The urease (Urs) was covalently immobilized onto an electrode made of gold nanoparticles functionalized with hyperbranched polyester-Boltron^® H40 (H40-Au) coated onto an indium-tin oxide (ITO) covered glass substrate. The covalent linkage between the Urs enzyme and H40-Au nanoparticles provided the resulting enzyme electrode (Urs/H40-Au/ITO) with a high level of enzyme immobilization and excellent lifetime stability. The response studies were carried out as a function of urea concentration with amperometric and photometric measurements. The biosensor based on Urs/H40-Au/ITO as the working electrode showed a linear current response to the urea concentration ranging from 0.01 to 35 mM. The urea biosensor exhibited a sensitivity of 7.48 nA/mM with a response time of 3 s. The Michaelis-Menten constant for the Urs/H40-Au/ITO biosensor was calculated to be 0.96 mM, indicating the Urs enzyme immobilized on the electrode surface had a high affinity to urea.

Original language	English (US)
Pages (from-to)	1401-1407
Number of pages	7
Journal	Talanta
Volume	78
Issue number	4-5
DOIs	https://doi.org/10.1016/j.talanta.2009.02.038
State	Published - Jun 15 2009

Keywords

Amperometric detection
Boltron H40
Covalent immobilization
Gold nanoparticles
Urea biosensor

ASJC Scopus subject areas

General Chemistry

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10.1016/j.talanta.2009.02.038

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title = "An amperometric urea biosensor based on covalently immobilized urease on an electrode made of hyperbranched polyester functionalized gold nanoparticles",

abstract = "An amperometric biosensor was fabricated for the quantitative determination of urea in aqueous medium using hematein, a pH-sensitive natural dye. The urease (Urs) was covalently immobilized onto an electrode made of gold nanoparticles functionalized with hyperbranched polyester-Boltron{\textregistered} H40 (H40-Au) coated onto an indium-tin oxide (ITO) covered glass substrate. The covalent linkage between the Urs enzyme and H40-Au nanoparticles provided the resulting enzyme electrode (Urs/H40-Au/ITO) with a high level of enzyme immobilization and excellent lifetime stability. The response studies were carried out as a function of urea concentration with amperometric and photometric measurements. The biosensor based on Urs/H40-Au/ITO as the working electrode showed a linear current response to the urea concentration ranging from 0.01 to 35 mM. The urea biosensor exhibited a sensitivity of 7.48 nA/mM with a response time of 3 s. The Michaelis-Menten constant for the Urs/H40-Au/ITO biosensor was calculated to be 0.96 mM, indicating the Urs enzyme immobilized on the electrode surface had a high affinity to urea.",

keywords = "Amperometric detection, Boltron H40, Covalent immobilization, Gold nanoparticles, Urea biosensor",

author = "Ashutosh Tiwari and Santosh Aryal and Srikanth Pilla and Shaoqin Gong",

note = "Funding Information: The authors would like to acknowledge the financial support to carry out this work from National Science Foundation (NSF), USA under the project CMMI-0734881.",

year = "2009",

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AU - Tiwari, Ashutosh

AU - Aryal, Santosh

AU - Pilla, Srikanth

AU - Gong, Shaoqin

N1 - Funding Information: The authors would like to acknowledge the financial support to carry out this work from National Science Foundation (NSF), USA under the project CMMI-0734881.

PY - 2009/6/15

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N2 - An amperometric biosensor was fabricated for the quantitative determination of urea in aqueous medium using hematein, a pH-sensitive natural dye. The urease (Urs) was covalently immobilized onto an electrode made of gold nanoparticles functionalized with hyperbranched polyester-Boltron® H40 (H40-Au) coated onto an indium-tin oxide (ITO) covered glass substrate. The covalent linkage between the Urs enzyme and H40-Au nanoparticles provided the resulting enzyme electrode (Urs/H40-Au/ITO) with a high level of enzyme immobilization and excellent lifetime stability. The response studies were carried out as a function of urea concentration with amperometric and photometric measurements. The biosensor based on Urs/H40-Au/ITO as the working electrode showed a linear current response to the urea concentration ranging from 0.01 to 35 mM. The urea biosensor exhibited a sensitivity of 7.48 nA/mM with a response time of 3 s. The Michaelis-Menten constant for the Urs/H40-Au/ITO biosensor was calculated to be 0.96 mM, indicating the Urs enzyme immobilized on the electrode surface had a high affinity to urea.

AB - An amperometric biosensor was fabricated for the quantitative determination of urea in aqueous medium using hematein, a pH-sensitive natural dye. The urease (Urs) was covalently immobilized onto an electrode made of gold nanoparticles functionalized with hyperbranched polyester-Boltron® H40 (H40-Au) coated onto an indium-tin oxide (ITO) covered glass substrate. The covalent linkage between the Urs enzyme and H40-Au nanoparticles provided the resulting enzyme electrode (Urs/H40-Au/ITO) with a high level of enzyme immobilization and excellent lifetime stability. The response studies were carried out as a function of urea concentration with amperometric and photometric measurements. The biosensor based on Urs/H40-Au/ITO as the working electrode showed a linear current response to the urea concentration ranging from 0.01 to 35 mM. The urea biosensor exhibited a sensitivity of 7.48 nA/mM with a response time of 3 s. The Michaelis-Menten constant for the Urs/H40-Au/ITO biosensor was calculated to be 0.96 mM, indicating the Urs enzyme immobilized on the electrode surface had a high affinity to urea.

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An amperometric urea biosensor based on covalently immobilized urease on an electrode made of hyperbranched polyester functionalized gold nanoparticles

Abstract

Keywords

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