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A chemiluminescence flow immunosensor based on a porous monolithic metacrylate and polyethylene composite disc modified with Protein G

Jain, Seema Rani; Borowska, Ewa; Davidsson, Richard LU ; Tudorache, Madalina LU ; Pontén, Einar and Emnéus, Jenny LU (2004) In Biosensors & Bioelectronics 19(8). p.795-803
Abstract
A generic, fast, sensitive and new type of flow immunosensor has been developed. The basis is a monolithic porous poly(glycidyl methacrylate-co-trimethylolpropane trimethacrylate) polymer disc modified with protein G, placed in a fountain type flow cell compartment, in close proximity to a photomultiplier tube (PMT). Analyte and HRP labelled analyte derivative (tracer) compete for anti-analyte antibody binding sites. The mixture is then injected into the flow immunosensor system where the formed analyte- and tracer-antibody complexes are trapped by the monolithic protein G disc. The amount of bound tracer, inversely related to the concentration of analyte in the sample, is determined in a second step by injection of luminol, p-iodophenol... (More)
A generic, fast, sensitive and new type of flow immunosensor has been developed. The basis is a monolithic porous poly(glycidyl methacrylate-co-trimethylolpropane trimethacrylate) polymer disc modified with protein G, placed in a fountain type flow cell compartment, in close proximity to a photomultiplier tube (PMT). Analyte and HRP labelled analyte derivative (tracer) compete for anti-analyte antibody binding sites. The mixture is then injected into the flow immunosensor system where the formed analyte- and tracer-antibody complexes are trapped by the monolithic protein G disc. The amount of bound tracer, inversely related to the concentration of analyte in the sample, is determined in a second step by injection of luminol, p-iodophenol and H2O2, generating enhanced chemiluminescence (CL) with horseradish peroxidase (HRP). A third and final step is need for regeneration of the protein G disc so that a new analysis cycle can take place. The performance of the disc immunosensor system was compared with a one step continuous flow injection immunoassay (FIIA) system, using the same reagents and a protein G column, in terms of assay sensitivity and influence of matrix effects from various water samples (millipore-, tap- and surface water). The detection limit for the analyte atrazine in PBS and surface water (SW) was 0.208±0.004 g l−1 (PBS) and 0.59±0.120 g l−1 (SW) for the FIIA and 0.033±0.003 g l−1 (PBS) and 0.038±0.003 g l−1 (SW) for the disc immunosensor. Statistical comparison of the two systems shows that the disc immunosensor results were significantly less influenced by the sample matrix, which is explained by the fact that the sample in the FIIA arrives simultaneously with the matrix to the detector, whereas these are separated in time in the disc immunosensor system. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Solid phase reactor, Protein G, Monolith, Immunosensor, Chemiluminescence, Atrazine
in
Biosensors & Bioelectronics
volume
19
issue
8
pages
795 - 803
publisher
Elsevier
external identifiers
  • wos:000189220900003
  • pmid:15128098
  • scopus:1042275724
ISSN
1873-4235
DOI
10.1016/j.bios.2003.08.008
language
English
LU publication?
yes
id
bd504d97-d6c5-490e-ac49-fe5c2d38b8f7 (old id 141276)
date added to LUP
2007-06-27 08:55:45
date last changed
2017-01-01 06:55:53
@article{bd504d97-d6c5-490e-ac49-fe5c2d38b8f7,
  abstract     = {A generic, fast, sensitive and new type of flow immunosensor has been developed. The basis is a monolithic porous poly(glycidyl methacrylate-co-trimethylolpropane trimethacrylate) polymer disc modified with protein G, placed in a fountain type flow cell compartment, in close proximity to a photomultiplier tube (PMT). Analyte and HRP labelled analyte derivative (tracer) compete for anti-analyte antibody binding sites. The mixture is then injected into the flow immunosensor system where the formed analyte- and tracer-antibody complexes are trapped by the monolithic protein G disc. The amount of bound tracer, inversely related to the concentration of analyte in the sample, is determined in a second step by injection of luminol, p-iodophenol and H2O2, generating enhanced chemiluminescence (CL) with horseradish peroxidase (HRP). A third and final step is need for regeneration of the protein G disc so that a new analysis cycle can take place. The performance of the disc immunosensor system was compared with a one step continuous flow injection immunoassay (FIIA) system, using the same reagents and a protein G column, in terms of assay sensitivity and influence of matrix effects from various water samples (millipore-, tap- and surface water). The detection limit for the analyte atrazine in PBS and surface water (SW) was 0.208±0.004 g l−1 (PBS) and 0.59±0.120 g l−1 (SW) for the FIIA and 0.033±0.003 g l−1 (PBS) and 0.038±0.003 g l−1 (SW) for the disc immunosensor. Statistical comparison of the two systems shows that the disc immunosensor results were significantly less influenced by the sample matrix, which is explained by the fact that the sample in the FIIA arrives simultaneously with the matrix to the detector, whereas these are separated in time in the disc immunosensor system.},
  author       = {Jain, Seema Rani and Borowska, Ewa and Davidsson, Richard and Tudorache, Madalina and Pontén, Einar and Emnéus, Jenny},
  issn         = {1873-4235},
  keyword      = {Solid phase reactor,Protein G,Monolith,Immunosensor,Chemiluminescence,Atrazine},
  language     = {eng},
  number       = {8},
  pages        = {795--803},
  publisher    = {Elsevier},
  series       = {Biosensors & Bioelectronics},
  title        = {A chemiluminescence flow immunosensor based on a porous monolithic metacrylate and polyethylene composite disc modified with Protein G},
  url          = {http://dx.doi.org/10.1016/j.bios.2003.08.008},
  volume       = {19},
  year         = {2004},
}