[Interaction between polysaccharides and interferon-gamma using an improved ELISA approach].
To establish an ELISA approach to study the interaction of polysaccharides with cytokine in vitro.



The heparin  BSA complexes (HBC) were synthesized with a chemistry method and separated using a 1 X 90 cm column of Separose 4B. After identification of the complex via SDS-PAGE,the wells of ELISA plates were coated with HBC and the interaction of HBC with interferon-gamma (IFN-gamma) was detected. The effects of heparin, low molecular weight heparin (LMW heparin), chondroitin sulfate (CS), hyaluronic acid (HA) and carrageenans on the binding of HBC to IFN-gamma were tested in this system.



Human recombinant IFN-gamma bound to heparin in a concentration  dependent manner, the binding of IFN-gamma to HBC was detected at the concentration of 0.25 ng, and saturated at around 2 ng. Free heparin, LMW heparin, CS,HA and carrageenans competed for the binding of IFN-gamma to HBC with significant different ability. The IC(50)concentrations of heparin and LMW heparin were 2.40 microg/ml and 18.60 microg/ml respectively.



IFN-gamma is a cytokine with high binding affinity to heparin and carrageenans family but poor to CS-A and CS-C. ELISA is a simple, sensitive approach to detect the interaction of polysaccharides with cytokine in vitro.
Department of Biochemistry and Molecular Biology, College of Medicine, Zhejiang University, Hangzhou 310031, China.
Wei-yun
WY
Feng
Lu-hang
LH
Zhao
Ke-yi
KY
Wang
D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006493:Heparin / Q000378:metabolism; D007371:Interferon-gamma / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism
D011134:Polysaccharides; D012710:Serum Albumin, Bovine; D007371:Interferon-gamma; D006493:Heparin