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Microfluidic enzyme immunoassay using silicon microchip with immobilized antibodies and chemiluminescence detection

Yakovleva, J; Davidsson, Richard LU ; Lobanova, A; Bengtsson, Martin LU ; Eremin, S; Laurell, Thomas LU and Emnéus, Jenny LU (2002) In Analytical Chemistry 74(13). p.2994-3004
Abstract
Silicon microchips with immobilized antibodies were used to develop microfluidic enzyme immunoassays using chemiluminescence detection and horseradish peroxidase (HRP) as the enzyme label. Polyclonal anti-atrazine antibodies were coupled to the silicon microchip surface with an overall dimension of 13.1 x 3.2 mm, comprising 42 porous flow channels of 235-mum depth and 25-mum width. Different immobilization protocols based on covalent or noncovalent modification of the silica surface with 3-aminopropyltriethoxysilane (APTES) or 3-glycidoxypropyltrimethoxysilane (GOPS), linear polyethylenimine (LPEI, MW 750 000), or branched polyethylenimine (BPEI, MW 25 000), followed by adsorption or covalent attachment of the antibody, were evaluated to... (More)
Silicon microchips with immobilized antibodies were used to develop microfluidic enzyme immunoassays using chemiluminescence detection and horseradish peroxidase (HRP) as the enzyme label. Polyclonal anti-atrazine antibodies were coupled to the silicon microchip surface with an overall dimension of 13.1 x 3.2 mm, comprising 42 porous flow channels of 235-mum depth and 25-mum width. Different immobilization protocols based on covalent or noncovalent modification of the silica surface with 3-aminopropyltriethoxysilane (APTES) or 3-glycidoxypropyltrimethoxysilane (GOPS), linear polyethylenimine (LPEI, MW 750 000), or branched polyethylenimine (BPEI, MW 25 000), followed by adsorption or covalent attachment of the antibody, were evaluated to reach the best reusability, stability, and sensitivity of the microfluidic enzyme immunoassay (muFEIA). Adsorption of antibodies on a LPEI-modified silica surface and covalent attachment to physically adsorbed BPEI lead to unstable antibody coatings. Covalent coupling of antibodies via glutaraldehyde (GA) to three different functionalized silica surfaces (APTES-GA, LPEI-GA, and GOPS-BPEI-GA) resulted in antibody coatings that could be completely regenerated using 0.4 M glycine/HCl, pH 2.2. The buffer composition was shown to have a dramatic effect on the assay stability, where the commonly used phosphate buffer saline was proved to be the least suitable choice. The best long-term stability was obtained for the LPEI-GA surface with no loss of antibody activity during one month. The detection limits in the muFEIA for the three different immuno surfaces were 45, 3.8, and 0.80 ng/L (209, 17.7, and 3.7 pM) for APTES-GA, LPEI-GA, and GOPS-BPEI-GA, respectively. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Analytical Chemistry
volume
74
issue
13
pages
2994 - 3004
publisher
The American Chemical Society
external identifiers
  • wos:000176589800021
  • pmid:12141657
  • scopus:0036646094
ISSN
1520-6882
DOI
10.1021/ac015645b
language
English
LU publication?
yes
id
7afcb356-cc72-4747-b989-79811b360f16 (old id 334234)
date added to LUP
2007-11-09 16:13:38
date last changed
2017-10-29 03:39:25
@article{7afcb356-cc72-4747-b989-79811b360f16,
  abstract     = {Silicon microchips with immobilized antibodies were used to develop microfluidic enzyme immunoassays using chemiluminescence detection and horseradish peroxidase (HRP) as the enzyme label. Polyclonal anti-atrazine antibodies were coupled to the silicon microchip surface with an overall dimension of 13.1 x 3.2 mm, comprising 42 porous flow channels of 235-mum depth and 25-mum width. Different immobilization protocols based on covalent or noncovalent modification of the silica surface with 3-aminopropyltriethoxysilane (APTES) or 3-glycidoxypropyltrimethoxysilane (GOPS), linear polyethylenimine (LPEI, MW 750 000), or branched polyethylenimine (BPEI, MW 25 000), followed by adsorption or covalent attachment of the antibody, were evaluated to reach the best reusability, stability, and sensitivity of the microfluidic enzyme immunoassay (muFEIA). Adsorption of antibodies on a LPEI-modified silica surface and covalent attachment to physically adsorbed BPEI lead to unstable antibody coatings. Covalent coupling of antibodies via glutaraldehyde (GA) to three different functionalized silica surfaces (APTES-GA, LPEI-GA, and GOPS-BPEI-GA) resulted in antibody coatings that could be completely regenerated using 0.4 M glycine/HCl, pH 2.2. The buffer composition was shown to have a dramatic effect on the assay stability, where the commonly used phosphate buffer saline was proved to be the least suitable choice. The best long-term stability was obtained for the LPEI-GA surface with no loss of antibody activity during one month. The detection limits in the muFEIA for the three different immuno surfaces were 45, 3.8, and 0.80 ng/L (209, 17.7, and 3.7 pM) for APTES-GA, LPEI-GA, and GOPS-BPEI-GA, respectively.},
  author       = {Yakovleva, J and Davidsson, Richard and Lobanova, A and Bengtsson, Martin and Eremin, S and Laurell, Thomas and Emnéus, Jenny},
  issn         = {1520-6882},
  language     = {eng},
  number       = {13},
  pages        = {2994--3004},
  publisher    = {The American Chemical Society},
  series       = {Analytical Chemistry},
  title        = {Microfluidic enzyme immunoassay using silicon microchip with immobilized antibodies and chemiluminescence detection},
  url          = {http://dx.doi.org/10.1021/ac015645b},
  volume       = {74},
  year         = {2002},
}