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

Jürgen Scheller, Marina Kovaleva, Björn Rabe, Jutta Eichler, Karl-Josef Kallen, Stefan Rose-John, Marina Kovaleva

Research output: Contribution to journalArticle

8 Citations (Scopus)

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.
Original languageEnglish
Pages (from-to)93-100
Number of pages8
JournalJournal of Immunological Methods
Volume291
Issue number1-2
DOIs
Publication statusPublished - Aug 2004

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Interleukin-6 Receptors
Interleukin-6
Monoclonal Antibodies
Enzymes
Cytokine Receptor gp130
Small Molecule Libraries
Suramin
Competitive Binding
Human Development
Peritonitis
Crohn Disease
Rheumatoid Arthritis
Homeostasis
Cytokines
Peptides
Membranes
Inhibition (Psychology)
Pharmaceutical Preparations

Keywords

  • Antibodies, Monoclonal
  • Antigens, CD
  • Binding, Competitive
  • Cell Line
  • Cytokine Receptor gp130
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Interleukin-6
  • Macromolecular Substances
  • Membrane Glycoproteins
  • Protein Binding
  • Receptors, Interleukin-6

Cite this

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. / Scheller, Jürgen; Kovaleva, Marina; Rabe, Björn; Eichler, Jutta; Kallen, Karl-Josef; Rose-John, Stefan; Kovaleva, Marina.

In: Journal of Immunological Methods, Vol. 291, No. 1-2, 08.2004, p. 93-100.

Research output: Contribution to journalArticle

Scheller, Jürgen ; Kovaleva, Marina ; Rabe, Björn ; Eichler, Jutta ; Kallen, Karl-Josef ; Rose-John, Stefan ; Kovaleva, Marina. / 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. In: Journal of Immunological Methods. 2004 ; Vol. 291, No. 1-2. pp. 93-100.
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AB - 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.

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