Development and validation of a multiplex fluorescent microsphere immunoassay assay for detection of porcine cytokines

SA Hall*, SH Ison, C Owles, J Coe, DA Sandercock, AJ Zanella

*Corresponding author for this work

Research output: Contribution to journalArticle

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Abstract

Cytokines are cell signalling proteins that mediate a number of different physiological responses. The accurate measurement of cytokine profiles is important for a variety of diagnostic and prognostic scenarios in relation to animal health and welfare. Simultaneous quantification of cytokine profiles in a single sample is now possible using fluorescent microsphere immunoassays (FMIA). We describe the development and validation of a novel multiplex assay using the Bio-Plex® 200 system to quantify cytokines in five different porcine tissues (brain, placenta, synovial tissue and fluid, plasma). The cytokine profiles are both tissue, and research hypothesis, -dependent but include Interleukin-1beta (IL-1β), Interleukin-4 (IL-4), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-10 (IL-10) and Tumor necrosis factor (TNF-α).

This methods paper is reported in two parts: the development of a FMIA for porcine tissues and validation of pre-treatment for optimal cytokine recovery in porcine brain, placenta, synovial tissue and plasma. Validation steps are critical in ensuring an assay is suitable for novel sample types.
Original languageEnglish
Pages (from-to)1218-1227
Number of pages10
JournalMethodsX
Volume6
Early online date17 May 2019
DOIs
Publication statusPrint publication - 2019

Fingerprint

Microspheres
Immunoassay
Assays
Swine
Cytokines
Tissue
Placenta
Brain
Cell signaling
Plasmas
Animal Welfare
Synovial Fluid
Interleukin-8
Interleukin-1beta
Interleukin-4
Interleukin-10
Interleukin-6
Animals
Tumor Necrosis Factor-alpha
Health

Keywords

  • Porcine
  • Cytokines
  • FMIA
  • Inflammatory
  • Pigs
  • Plasma
  • Synovial tissue
  • Synovial fluid
  • Brain
  • Placenta
  • BioPlex

Cite this

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title = "Development and validation of a multiplex fluorescent microsphere immunoassay assay for detection of porcine cytokines",
abstract = "Cytokines are cell signalling proteins that mediate a number of different physiological responses. The accurate measurement of cytokine profiles is important for a variety of diagnostic and prognostic scenarios in relation to animal health and welfare. Simultaneous quantification of cytokine profiles in a single sample is now possible using fluorescent microsphere immunoassays (FMIA). We describe the development and validation of a novel multiplex assay using the Bio-Plex{\circledR} 200 system to quantify cytokines in five different porcine tissues (brain, placenta, synovial tissue and fluid, plasma). The cytokine profiles are both tissue, and research hypothesis, -dependent but include Interleukin-1beta (IL-1β), Interleukin-4 (IL-4), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-10 (IL-10) and Tumor necrosis factor (TNF-α).This methods paper is reported in two parts: the development of a FMIA for porcine tissues and validation of pre-treatment for optimal cytokine recovery in porcine brain, placenta, synovial tissue and plasma. Validation steps are critical in ensuring an assay is suitable for novel sample types.",
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Development and validation of a multiplex fluorescent microsphere immunoassay assay for detection of porcine cytokines. / Hall, SA; Ison, SH; Owles, C; Coe, J; Sandercock, DA; Zanella, AJ.

In: MethodsX, Vol. 6, 2019, p. 1218-1227.

Research output: Contribution to journalArticle

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AU - Ison, SH

AU - Owles, C

AU - Coe, J

AU - Sandercock, DA

AU - Zanella, AJ

PY - 2019

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AB - Cytokines are cell signalling proteins that mediate a number of different physiological responses. The accurate measurement of cytokine profiles is important for a variety of diagnostic and prognostic scenarios in relation to animal health and welfare. Simultaneous quantification of cytokine profiles in a single sample is now possible using fluorescent microsphere immunoassays (FMIA). We describe the development and validation of a novel multiplex assay using the Bio-Plex® 200 system to quantify cytokines in five different porcine tissues (brain, placenta, synovial tissue and fluid, plasma). The cytokine profiles are both tissue, and research hypothesis, -dependent but include Interleukin-1beta (IL-1β), Interleukin-4 (IL-4), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-10 (IL-10) and Tumor necrosis factor (TNF-α).This methods paper is reported in two parts: the development of a FMIA for porcine tissues and validation of pre-treatment for optimal cytokine recovery in porcine brain, placenta, synovial tissue and plasma. Validation steps are critical in ensuring an assay is suitable for novel sample types.

KW - Porcine

KW - Cytokines

KW - FMIA

KW - Inflammatory

KW - Pigs

KW - Plasma

KW - Synovial tissue

KW - Synovial fluid

KW - Brain

KW - Placenta

KW - BioPlex

U2 - 10.1016/j.mex.2019.05.013

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