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 statusFirst published - 17 May 2019

Keywords

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

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  • Profiles

    No photo of Sarah Hall

    Sarah Hall

    Person: Academic contract that is research only

    No photo of Dale Sandercock

    Dale Sandercock

    Person: Academic contract that is research only

    Equipment

    Biomarkers Laboratory

    Arianne Lowe (Manager)

    Veterinary And Animal Sciences

    Facility/equipment: Facility

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