Development of a monoclonal antibody-based enzyme-linked immunoabsorbent assay for the binding of gp130 to the IL-6/IL-6R complex and its competitive inhibition

J Immunol Methods. 2004 Aug;291(1-2):93-100. doi: 10.1016/j.jim.2004.05.002.

Abstract

The proinflammatory cytokine IL-6 binds to the membrane bound or soluble IL-6 receptor (IL-6R) and activates an intracellular signaling cascade after complex formation with two gp130 molecules. These mediate general homeostasis and orchestrates the immune response during disease. Trans-signalling via the soluble IL-6R has importance for the development and maintenance of human diseases like Crohn's disease, peritonitis and rheumatoid arthritis. We have developed an enzyme-linked immunoabsorbent assay (ELISA) that detects the binding of gp130 to the IL-6/sIL-6R complex. Competitive binding of sgp130-Fc, viral IL-6, and the inhibitory drug Suramin to gp130 has been demonstrated. The assay can be used for high-throughput screening of peptide and chemical compound libraries for the identification of new agonists and antagonists of the binding between gp130 and IL-6/sIL-6R.

MeSH terms

  • Antibodies, Monoclonal / immunology*
  • Antigens, CD / analysis*
  • Antigens, CD / immunology*
  • Binding, Competitive
  • Cell Line
  • Cytokine Receptor gp130
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Humans
  • Interleukin-6 / antagonists & inhibitors
  • Interleukin-6 / immunology*
  • Macromolecular Substances
  • Membrane Glycoproteins / analysis*
  • Membrane Glycoproteins / antagonists & inhibitors
  • Membrane Glycoproteins / immunology*
  • Protein Binding
  • Receptors, Interleukin-6 / antagonists & inhibitors
  • Receptors, Interleukin-6 / immunology*

Substances

  • Antibodies, Monoclonal
  • Antigens, CD
  • IL6ST protein, human
  • Interleukin-6
  • Macromolecular Substances
  • Membrane Glycoproteins
  • Receptors, Interleukin-6
  • Cytokine Receptor gp130