Interaction of polysaccharides with interferon-gamma using an improved ELISA approach

摘要

Establishes an ELISA approach to study the interaction of polysaccharides with cytokine in vitro.Synthesizes and separates heparin-BSA complex,after identification of the complex via SDS-PAGE,the ELISA plate wells are coated with HBC,detecting the interaction of interferon-gamma (IFN-y) with heparin-BSA complex.We have investigated recombinant human IFN-gamma (rhTFN-gamma) binds to heparin in a concentration-dependent manner.Free heparin and low molecular weight heparin (LMW heparin) competes for the binding of IFN-gamma to heparin,the IC_(50) concentrations give 2.40 and 18.60 ng/ml,respectively.Other polysaccharides either with sulfate group or without sulfate group compete for the binding of IFN-gamma to HBC with significant different ability.The results suggest that IFN-gamma is a cytokine binding to some polysaccharides including heparin,chondroitin sulfate,fucoidan and carrageenans family with different affinity.ELISA is a simple,sensitive approach to detect the interaction of cytokine with polysaccharides.