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Development of fluorescence based flow immunoassays utilising restricted access columns

Önnerfjord, P. LU orcid and Marko-Varga, G. LU (2000) In Chromatographia 51(3-4). p.199-204
Abstract

The development of high-speed flow immunoassay techniques is described. The principles are based on heterogeneous flow immunoassay interactions. High sample throughput can be used for screening small analytes in a number of biological matrices originating from samples of water from environmentally polluted areas, or biological fluids such as urine and plasma. The immunochemical detection principle is based on chromatographic separation of the immunocomplex formed (AbAg or AbAg*) and the free antigen (Ag) by a restricted access (RA) column, utilising size-exclusion and reversed phase mechanisms. A fluorescein-labelled analyte (Ag*) was used in the competitive assay format with fluorescence detection. Sample... (More)

The development of high-speed flow immunoassay techniques is described. The principles are based on heterogeneous flow immunoassay interactions. High sample throughput can be used for screening small analytes in a number of biological matrices originating from samples of water from environmentally polluted areas, or biological fluids such as urine and plasma. The immunochemical detection principle is based on chromatographic separation of the immunocomplex formed (AbAg or AbAg*) and the free antigen (Ag) by a restricted access (RA) column, utilising size-exclusion and reversed phase mechanisms. A fluorescein-labelled analyte (Ag*) was used in the competitive assay format with fluorescence detection. Sample throughput was 80 h-1 and detection limits 1.4 nM (300pgml-1) for atrazine and 2.3 nM (500 pg ml-1) for the sum of triazines. Analyses could be performed at a sample throughput of 400 6 h-1 shift. Basic immunoaffinity interactions of a number of immunoreagents, using fluorescence polarisation were studied and outlined both for triazines and for 2,4-D. Structural variations in tracer synthesis confirmed that this is an important part in the design and optimisation of flow immuno methodologies.

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author
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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Fluorescence flow immunoassay, Fluorescence polarisation, Restricted access material, Triazines
in
Chromatographia
volume
51
issue
3-4
pages
6 pages
publisher
Vieweg Verlag
external identifiers
  • scopus:0034012905
ISSN
0009-5893
DOI
10.1007/BF02490565
language
English
LU publication?
yes
id
29a1b5e2-3bcf-4d04-a682-9459b46c8965
date added to LUP
2016-10-14 11:35:26
date last changed
2022-01-30 06:45:44
@article{29a1b5e2-3bcf-4d04-a682-9459b46c8965,
  abstract     = {{<p>The development of high-speed flow immunoassay techniques is described. The principles are based on heterogeneous flow immunoassay interactions. High sample throughput can be used for screening small analytes in a number of biological matrices originating from samples of water from environmentally polluted areas, or biological fluids such as urine and plasma. The immunochemical detection principle is based on chromatographic separation of the immunocomplex formed (AbAg or AbAg<sup>*</sup>) and the free antigen (Ag) by a restricted access (RA) column, utilising size-exclusion and reversed phase mechanisms. A fluorescein-labelled analyte (Ag<sup>*</sup>) was used in the competitive assay format with fluorescence detection. Sample throughput was 80 h<sup>-1</sup> and detection limits 1.4 nM (300pgml<sup>-1</sup>) for atrazine and 2.3 nM (500 pg ml<sup>-1</sup>) for the sum of triazines. Analyses could be performed at a sample throughput of 400 6 h<sup>-1</sup> shift. Basic immunoaffinity interactions of a number of immunoreagents, using fluorescence polarisation were studied and outlined both for triazines and for 2,4-D. Structural variations in tracer synthesis confirmed that this is an important part in the design and optimisation of flow immuno methodologies.</p>}},
  author       = {{Önnerfjord, P. and Marko-Varga, G.}},
  issn         = {{0009-5893}},
  keywords     = {{Fluorescence flow immunoassay; Fluorescence polarisation; Restricted access material; Triazines}},
  language     = {{eng}},
  number       = {{3-4}},
  pages        = {{199--204}},
  publisher    = {{Vieweg Verlag}},
  series       = {{Chromatographia}},
  title        = {{Development of fluorescence based flow immunoassays utilising restricted access columns}},
  url          = {{http://dx.doi.org/10.1007/BF02490565}},
  doi          = {{10.1007/BF02490565}},
  volume       = {{51}},
  year         = {{2000}},
}