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.
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.
Scheller
Jürgen
J
Institut für Biochemie, Christian-Albrechts Universität zu Kiel, Olshausenstr.40, D-24098 Kiel, Germany. jscheller@biochem.uni-kiel.de
Kovaleva
Marina
M
Rabe
Björn
B
Eichler
Jutta
J
Kallen
Karl-Josef
KJ
Rose-John
Stefan
S
D000911:Antibodies, Monoclonal / Q000276:immunology*; D015703:Antigens, CD / Q000032:analysis* / Q000276:immunology*; D001667:Binding, Competitive; D002460:Cell Line; D050822:Cytokine Receptor gp130; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D015850:Interleukin-6 / Q000276:immunology*; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000276:immunology*; D011485:Protein Binding; D019947:Receptors, Interleukin-6 / Q000037:antagonists & inhibitors / Q000276:immunology*; D000093242:Interleukin-6 Inhibitors
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D050822:Cytokine Receptor gp130; D015850:Interleukin-6; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D019947:Receptors, Interleukin-6; D000093242:Interleukin-6 Inhibitors; C494199:IL6ST protein, human