Mette Bjerre

Simultaneous detection of porcine cytokines by multiplex analysis: Development of magnetic bioplex assay

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Simultaneous detection of porcine cytokines by multiplex analysis: Development of magnetic bioplex assay. / Bjerre, Mette; Hansen, Troels Krarup; Flyvbjerg, Allan; Tønnesen, Else.

I: Veterinary Immunology and Immunopathology, Bind 130, Nr. 1-2, 2009, s. 53-8.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

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Bjerre, Mette ; Hansen, Troels Krarup ; Flyvbjerg, Allan ; Tønnesen, Else. / Simultaneous detection of porcine cytokines by multiplex analysis: Development of magnetic bioplex assay. I: Veterinary Immunology and Immunopathology. 2009 ; Bind 130, Nr. 1-2. s. 53-8.

Bibtex

@article{c221f3a04b5211de8dc9000ea68e967b,
title = "Simultaneous detection of porcine cytokines by multiplex analysis: Development of magnetic bioplex assay",
abstract = "Multiplex assays for analysis of human and rodent cytokines are highly developed, while development of porcine cytokine assays needs further attention. In order to follow the cytokine response in consecutive porcine samples, in which the sample volume may be limited, we have developed multiplex immunoassays for simultaneous detection of porcine cytokines interleukin (IL)-1beta, IL-6, IL-8, IL-10, tumour necrosis factor alpha (TNF-alpha), and heat shock protein 32 (Hsp32). Antibodies against porcine cytokines were coupled to magnetic microspheres. Quantification was obtained with biotinylated antibodies followed by PE-labelled streptavidin and measurements by Luminex(100). Validation and cross-reaction experiments revealed detection limits below 5-20ng/L, recovery of recombinant cytokines in spiked plasma between 80 and 110%, and intra- and inter-assay variation between 5 and 15%. No cross-reaction between assays was found. However, for optimal sensitivity the assays were performed as a 2-plex (IL-1beta and Hsp32) and a 4-plex (IL-6, IL-8, IL-10, and TNF-alpha). Cytokine levels were determined in plasma samples from a porcine model of acute endotoxaemia and the levels correlated to previously published concentrations.",
author = "Mette Bjerre and Hansen, {Troels Krarup} and Allan Flyvbjerg and Else T{\o}nnesen",
year = "2009",
doi = "10.1016/j.vetimm.2009.01.007",
language = "English",
volume = "130",
pages = "53--8",
journal = "Veterinary Immunology and Immunopathology",
issn = "0165-2427",
publisher = "Elsevier BV",
number = "1-2",

}

RIS

TY - JOUR

T1 - Simultaneous detection of porcine cytokines by multiplex analysis: Development of magnetic bioplex assay

AU - Bjerre, Mette

AU - Hansen, Troels Krarup

AU - Flyvbjerg, Allan

AU - Tønnesen, Else

PY - 2009

Y1 - 2009

N2 - Multiplex assays for analysis of human and rodent cytokines are highly developed, while development of porcine cytokine assays needs further attention. In order to follow the cytokine response in consecutive porcine samples, in which the sample volume may be limited, we have developed multiplex immunoassays for simultaneous detection of porcine cytokines interleukin (IL)-1beta, IL-6, IL-8, IL-10, tumour necrosis factor alpha (TNF-alpha), and heat shock protein 32 (Hsp32). Antibodies against porcine cytokines were coupled to magnetic microspheres. Quantification was obtained with biotinylated antibodies followed by PE-labelled streptavidin and measurements by Luminex(100). Validation and cross-reaction experiments revealed detection limits below 5-20ng/L, recovery of recombinant cytokines in spiked plasma between 80 and 110%, and intra- and inter-assay variation between 5 and 15%. No cross-reaction between assays was found. However, for optimal sensitivity the assays were performed as a 2-plex (IL-1beta and Hsp32) and a 4-plex (IL-6, IL-8, IL-10, and TNF-alpha). Cytokine levels were determined in plasma samples from a porcine model of acute endotoxaemia and the levels correlated to previously published concentrations.

AB - Multiplex assays for analysis of human and rodent cytokines are highly developed, while development of porcine cytokine assays needs further attention. In order to follow the cytokine response in consecutive porcine samples, in which the sample volume may be limited, we have developed multiplex immunoassays for simultaneous detection of porcine cytokines interleukin (IL)-1beta, IL-6, IL-8, IL-10, tumour necrosis factor alpha (TNF-alpha), and heat shock protein 32 (Hsp32). Antibodies against porcine cytokines were coupled to magnetic microspheres. Quantification was obtained with biotinylated antibodies followed by PE-labelled streptavidin and measurements by Luminex(100). Validation and cross-reaction experiments revealed detection limits below 5-20ng/L, recovery of recombinant cytokines in spiked plasma between 80 and 110%, and intra- and inter-assay variation between 5 and 15%. No cross-reaction between assays was found. However, for optimal sensitivity the assays were performed as a 2-plex (IL-1beta and Hsp32) and a 4-plex (IL-6, IL-8, IL-10, and TNF-alpha). Cytokine levels were determined in plasma samples from a porcine model of acute endotoxaemia and the levels correlated to previously published concentrations.

U2 - 10.1016/j.vetimm.2009.01.007

DO - 10.1016/j.vetimm.2009.01.007

M3 - Journal article

C2 - 19230983

VL - 130

SP - 53

EP - 58

JO - Veterinary Immunology and Immunopathology

JF - Veterinary Immunology and Immunopathology

SN - 0165-2427

IS - 1-2

ER -