Multiplex Fluorescent Bead-Based Immunoassay for the Detection of Cytokines, Chemokines, and Growth Factors

Methods Mol Biol. 2021:2261:247-262. doi: 10.1007/978-1-0716-1186-9_15.

Abstract

The comprehensive analysis of serum cytokine levels can be challenging due to low sample volumes and time consuming when using single-target methods like enzyme-linked immunosorbent assay (ELISA). Bead-based detection systems allow the simultaneous detection of multiple analytes using minimal sample volumes. Here we describe the use of a multiplex cytokine, chemokine, and growth factor assay for mouse cytokines in a 96-well format. This assay is based on antibody-coupled fluorescent magnetic beads combined with biotinylated secondary detection antibody followed by fluorescent-tagged streptavidin in a sandwich-like composition. Final assay readout provides the concentrations of 23 different cytokines, chemokines, and growth factors in up to 76 samples.

Keywords: Bead-based; Bioplex; Chemokine; Cytokine; Endotoxemia; Glucocorticoid receptor; Inflammation; Intensive care; Multiplex; Serum.

MeSH terms

  • Animals
  • Biotinylation
  • Chemokines / blood*
  • Cytokines / blood*
  • Disease Models, Animal
  • Endotoxemia / blood*
  • Endotoxemia / chemically induced
  • Endotoxemia / genetics
  • Fluorescent Antibody Technique, Indirect*
  • Fluoroimmunoassay*
  • Intercellular Signaling Peptides and Proteins / blood*
  • Lipopolysaccharides
  • Mice
  • Mice, Knockout
  • Proteomics*
  • Receptors, Glucocorticoid / deficiency
  • Receptors, Glucocorticoid / genetics

Substances

  • Chemokines
  • Cytokines
  • Intercellular Signaling Peptides and Proteins
  • Lipopolysaccharides
  • Receptors, Glucocorticoid