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Microfluidic biosensing systems - Part I. Development and optimisation of enzymatic chemiluminescent mu-biosensors based on silicon microchips

Davidsson, Richard LU ; Genin, F; Bengtsson, Martin LU ; Laurell, Thomas LU and Emnéus, Jenny LU (2004) In Lab on A Chip 4(5). p.481-487
Abstract
Chemiluminescent (CL) enzyme-based flow-through microchip biosensors (mu-biosensors) for detection of glucose and ethanol were developed for the purpose of monitoring real-time production and release of glucose and ethanol from microchip immobilised yeast cells. Part I of this study focuses on the development and optimisation of the mu-biosensors in a microfluidic sequential injection analysis (muSIA) system. Glucose oxidase (GOX) or alcohol oxidase (AOX) was co-immobilised with horseradish peroxidase (HRP) on porous silicon flow through microchips. The hydrogen peroxide ;produced from oxidation of the corresponding analyte (glucose or ethanol) took part in the chemiluminescent (CL) oxidation of luminol catalysed by HRP enhanced by... (More)
Chemiluminescent (CL) enzyme-based flow-through microchip biosensors (mu-biosensors) for detection of glucose and ethanol were developed for the purpose of monitoring real-time production and release of glucose and ethanol from microchip immobilised yeast cells. Part I of this study focuses on the development and optimisation of the mu-biosensors in a microfluidic sequential injection analysis (muSIA) system. Glucose oxidase (GOX) or alcohol oxidase (AOX) was co-immobilised with horseradish peroxidase (HRP) on porous silicon flow through microchips. The hydrogen peroxide ;produced from oxidation of the corresponding analyte (glucose or ethanol) took part in the chemiluminescent (CL) oxidation of luminol catalysed by HRP enhanced by addition of p-iodophenol ( PIP). All steps in the mSIA system, including control of syringe pump, multiposition valve (MPV) and data readout, were computer controlled. The influence of flow rate and luminol- and PIP concentration were investigated using a 2(3)-factor experiment using the GOX-HRP sensor. It was found that all estimated single factors and the highest order of interaction were significant. The optimum was found at 250 muM luminol and 150 muM PIP at a flow rate of 18 mul min(-1), the latter as a compromise between signal intensity and analysis time. Using the optimised system settings one sample was processed within 5 min. Two different immobilisation chemistries were investigated for both m-biosensors based on 3-aminopropyltriethoxsilane (APTS)- or polyethylenimine (PEI) functionalisation followed by glutaraldehyde (GA) activation. GOX-HRP mu-biosensors responded linear in a log-log format within the range 10-1000 mM glucose. Both had an operational stability of at least 8 days, but the PEI-GOX-HRP sensor was more sensitive. The AOX-HRP mu-biosensors responded linear (log-log) in the range between 1 and 10 mM ethanol, but the PEI-AOX-HRP sensor was in general more sensitive. Both sensors had an operational stability of at least 8 h, but with a half-life of 2-3 days. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Lab on A Chip
volume
4
issue
5
pages
481 - 487
publisher
Royal Society of Chemistry
external identifiers
  • wos:000224475200012
  • pmid:15472732
  • scopus:7944223971
ISSN
1473-0189
DOI
10.1039/b400894d
language
English
LU publication?
yes
id
a22dc166-a332-4e10-abe2-3d90c17ac964 (old id 138365)
date added to LUP
2007-06-26 13:56:03
date last changed
2017-08-06 03:31:49
@article{a22dc166-a332-4e10-abe2-3d90c17ac964,
  abstract     = {Chemiluminescent (CL) enzyme-based flow-through microchip biosensors (mu-biosensors) for detection of glucose and ethanol were developed for the purpose of monitoring real-time production and release of glucose and ethanol from microchip immobilised yeast cells. Part I of this study focuses on the development and optimisation of the mu-biosensors in a microfluidic sequential injection analysis (muSIA) system. Glucose oxidase (GOX) or alcohol oxidase (AOX) was co-immobilised with horseradish peroxidase (HRP) on porous silicon flow through microchips. The hydrogen peroxide ;produced from oxidation of the corresponding analyte (glucose or ethanol) took part in the chemiluminescent (CL) oxidation of luminol catalysed by HRP enhanced by addition of p-iodophenol ( PIP). All steps in the mSIA system, including control of syringe pump, multiposition valve (MPV) and data readout, were computer controlled. The influence of flow rate and luminol- and PIP concentration were investigated using a 2(3)-factor experiment using the GOX-HRP sensor. It was found that all estimated single factors and the highest order of interaction were significant. The optimum was found at 250 muM luminol and 150 muM PIP at a flow rate of 18 mul min(-1), the latter as a compromise between signal intensity and analysis time. Using the optimised system settings one sample was processed within 5 min. Two different immobilisation chemistries were investigated for both m-biosensors based on 3-aminopropyltriethoxsilane (APTS)- or polyethylenimine (PEI) functionalisation followed by glutaraldehyde (GA) activation. GOX-HRP mu-biosensors responded linear in a log-log format within the range 10-1000 mM glucose. Both had an operational stability of at least 8 days, but the PEI-GOX-HRP sensor was more sensitive. The AOX-HRP mu-biosensors responded linear (log-log) in the range between 1 and 10 mM ethanol, but the PEI-AOX-HRP sensor was in general more sensitive. Both sensors had an operational stability of at least 8 h, but with a half-life of 2-3 days.},
  author       = {Davidsson, Richard and Genin, F and Bengtsson, Martin and Laurell, Thomas and Emnéus, Jenny},
  issn         = {1473-0189},
  language     = {eng},
  number       = {5},
  pages        = {481--487},
  publisher    = {Royal Society of Chemistry},
  series       = {Lab on A Chip},
  title        = {Microfluidic biosensing systems - Part I. Development and optimisation of enzymatic chemiluminescent mu-biosensors based on silicon microchips},
  url          = {http://dx.doi.org/10.1039/b400894d},
  volume       = {4},
  year         = {2004},
}