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Flow immunochemical bio-recognition detection for the determination of interleukin-10 in cell samples

Kjellström, S LU ; Emnéus, J LU and Marko-Varga, G LU (2000) In Journal of Immunological Methods 246(1-2). p.30-119
Abstract

On- and off-line heterogeneous non-competitive flow immunoassays for the determination of Interleukin-10 are described. The sample containing IL-10 is mixed, either on-line in a reaction coil or off-line in a test tube, with fluorescent labelled anti-IL-10 antibodies to form an antibody-antigen complex. The labelled unbound antibodies are trapped on an immobilized IL-10 column whereas the IL-10-antibody complexes are eluted and detected downstream by a fluorescence detector. The optimization of the systems was performed with respect to choice of affinity support, flow rate, carrier buffer additives, pH and antibody-antigen association. Both bio recognition assays were tested with a spiked cell medium and the IL-10 detection limits in... (More)

On- and off-line heterogeneous non-competitive flow immunoassays for the determination of Interleukin-10 are described. The sample containing IL-10 is mixed, either on-line in a reaction coil or off-line in a test tube, with fluorescent labelled anti-IL-10 antibodies to form an antibody-antigen complex. The labelled unbound antibodies are trapped on an immobilized IL-10 column whereas the IL-10-antibody complexes are eluted and detected downstream by a fluorescence detector. The optimization of the systems was performed with respect to choice of affinity support, flow rate, carrier buffer additives, pH and antibody-antigen association. Both bio recognition assays were tested with a spiked cell medium and the IL-10 detection limits in this matrix was found to be 8 fmol using the off-line incubation mode and 40 fmol using the on-line incubation mode. The sample through-put was 26 and 40 samples per hour in the on-line and off-line incubation modes, respectively. IL-10 identification in the sample fractions was achieved using MALDI-TOF MS.

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Contribution to journal
publication status
published
subject
keywords
Antigen-Antibody Reactions, Chromatography, Affinity, Cross Reactions, Fluoroimmunoassay/methods, Humans, Interleukin-10/analysis, Kinetics, Recombinant Proteins/analysis, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Substrate Specificity
in
Journal of Immunological Methods
volume
246
issue
1-2
pages
30 - 119
publisher
Elsevier
external identifiers
  • scopus:0034564462
ISSN
0022-1759
DOI
10.1016/S0022-1759(00)00307-0
language
English
LU publication?
yes
id
ecd0e41d-93b0-45b0-b9e5-b299d0b9d5fc
date added to LUP
2018-06-05 09:04:44
date last changed
2018-06-10 05:29:17
@article{ecd0e41d-93b0-45b0-b9e5-b299d0b9d5fc,
  abstract     = {<p>On- and off-line heterogeneous non-competitive flow immunoassays for the determination of Interleukin-10 are described. The sample containing IL-10 is mixed, either on-line in a reaction coil or off-line in a test tube, with fluorescent labelled anti-IL-10 antibodies to form an antibody-antigen complex. The labelled unbound antibodies are trapped on an immobilized IL-10 column whereas the IL-10-antibody complexes are eluted and detected downstream by a fluorescence detector. The optimization of the systems was performed with respect to choice of affinity support, flow rate, carrier buffer additives, pH and antibody-antigen association. Both bio recognition assays were tested with a spiked cell medium and the IL-10 detection limits in this matrix was found to be 8 fmol using the off-line incubation mode and 40 fmol using the on-line incubation mode. The sample through-put was 26 and 40 samples per hour in the on-line and off-line incubation modes, respectively. IL-10 identification in the sample fractions was achieved using MALDI-TOF MS.</p>},
  author       = {Kjellström, S and Emnéus, J and Marko-Varga, G},
  issn         = {0022-1759},
  keyword      = {Antigen-Antibody Reactions,Chromatography, Affinity,Cross Reactions,Fluoroimmunoassay/methods,Humans,Interleukin-10/analysis,Kinetics,Recombinant Proteins/analysis,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization,Substrate Specificity},
  language     = {eng},
  month        = {12},
  number       = {1-2},
  pages        = {30--119},
  publisher    = {Elsevier},
  series       = {Journal of Immunological Methods},
  title        = {Flow immunochemical bio-recognition detection for the determination of interleukin-10 in cell samples},
  url          = {http://dx.doi.org/10.1016/S0022-1759(00)00307-0},
  volume       = {246},
  year         = {2000},
}