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High sample throughput flow immunoassay utilising restricted access columns for the separation of bound and free label

Önnerfjord, Patrik LU ; Eremin, Sergei A.; Emnéus, Jenny LU and Marko-Varga, György LU (1998) In Journal of Chromatography A 800(2). p.219-230
Abstract

A flow immunodetection system with high sample throughput capacity is described for the screening of various analytes. The immunochemical detection principle is based on the chromatographic separation of the formed immunocomplex (AbAg or AbAg*) and the free antigen (Ag) by a restricted access (RA) column, utilising size-exclusion and reversed-phase mechanism. A fluorescein labelled analyte (Ag*) was used in the competitive assay format with fluorescence detection. The speed and simplicity of the assay were the greatest advantages, allowing measurement of the analyte to be carried out in less than 1 min. The biocompatibility and capacity of the restricted access material allowed multiple injections of up to 5000, without any breakthrough... (More)

A flow immunodetection system with high sample throughput capacity is described for the screening of various analytes. The immunochemical detection principle is based on the chromatographic separation of the formed immunocomplex (AbAg or AbAg*) and the free antigen (Ag) by a restricted access (RA) column, utilising size-exclusion and reversed-phase mechanism. A fluorescein labelled analyte (Ag*) was used in the competitive assay format with fluorescence detection. The speed and simplicity of the assay were the greatest advantages, allowing measurement of the analyte to be carried out in less than 1 min. The biocompatibility and capacity of the restricted access material allowed multiple injections of up to 5000, without any breakthrough of the fluorescent tracer molecule and thus need for regeneration. The flow immunoassay was developed using the well-known atrazine herbicide and some transformation products as model compounds, due to their human toxicity and widespread use. The sample throughput was 80 samples per hour and the detection limits were 1.4 nM (300 pg/ml) for atrazine (Ab I) and 2.3 nM (500 pg/ml) for the sum of triazines (Ab II-III). Different sample matrices, PBS buffer, creek water, and urine were successfully applied in the flow system without the need for any sample handling step. For plasma samples an additional clean-up step using solid-phase extraction had to be included. The resulting detection limits for atrazine in plasma and water samples using this clean-up and trace enrichment procedure were found to be 2 ng/ml and 20 pg/ml, respectively. The analysis could be performed at a sample throughput rate of 400 per 6-h working shift.

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Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
keywords
Atrazine, Flow immunoassay, Pesticides, Restricted access column
in
Journal of Chromatography A
volume
800
issue
2
pages
219 - 230
publisher
Elsevier
external identifiers
  • Scopus:0032571498
ISSN
0021-9673
DOI
10.1016/S0021-9673(97)01159-X
language
English
LU publication?
yes
id
735a8c5b-26a2-4f42-8048-b52f28e93f55
date added to LUP
2016-10-14 11:38:21
date last changed
2016-11-28 16:17:35
@misc{735a8c5b-26a2-4f42-8048-b52f28e93f55,
  abstract     = {<p>A flow immunodetection system with high sample throughput capacity is described for the screening of various analytes. The immunochemical detection principle is based on the chromatographic separation of the formed immunocomplex (AbAg or AbAg*) and the free antigen (Ag) by a restricted access (RA) column, utilising size-exclusion and reversed-phase mechanism. A fluorescein labelled analyte (Ag*) was used in the competitive assay format with fluorescence detection. The speed and simplicity of the assay were the greatest advantages, allowing measurement of the analyte to be carried out in less than 1 min. The biocompatibility and capacity of the restricted access material allowed multiple injections of up to 5000, without any breakthrough of the fluorescent tracer molecule and thus need for regeneration. The flow immunoassay was developed using the well-known atrazine herbicide and some transformation products as model compounds, due to their human toxicity and widespread use. The sample throughput was 80 samples per hour and the detection limits were 1.4 nM (300 pg/ml) for atrazine (Ab I) and 2.3 nM (500 pg/ml) for the sum of triazines (Ab II-III). Different sample matrices, PBS buffer, creek water, and urine were successfully applied in the flow system without the need for any sample handling step. For plasma samples an additional clean-up step using solid-phase extraction had to be included. The resulting detection limits for atrazine in plasma and water samples using this clean-up and trace enrichment procedure were found to be 2 ng/ml and 20 pg/ml, respectively. The analysis could be performed at a sample throughput rate of 400 per 6-h working shift.</p>},
  author       = {Önnerfjord, Patrik and Eremin, Sergei A. and Emnéus, Jenny and Marko-Varga, György},
  issn         = {0021-9673},
  keyword      = {Atrazine,Flow immunoassay,Pesticides,Restricted access column},
  language     = {eng},
  month        = {03},
  number       = {2},
  pages        = {219--230},
  publisher    = {ARRAY(0xa12ed08)},
  series       = {Journal of Chromatography A},
  title        = {High sample throughput flow immunoassay utilising restricted access columns for the separation of bound and free label},
  url          = {http://dx.doi.org/10.1016/S0021-9673(97)01159-X},
  volume       = {800},
  year         = {1998},
}