A surface plasmon resonance (SPR) based biosensor has been used for studying the interaction of recombinant @entity1 @entity2841 ( @entity2841 ) with genetically engineered @entity7722 subunits of its specific receptor ( @entity10734 ). Western blot analysis of @entity10734 confirmed the correct size (80 kDa) and reactivity of these proteins against anti- @entity10734 polyclonal or monoclonal antibodies. @entity2841 was immobilized, using standard @entity6115 coupling methods, to the dextran-modified gold biosensor surface in order to capture @entity10734 subsequently injected over the sensing layer. @entity10734 were shown to specifically form complexes with the immobilized ligand. Pre-incubation of constant amounts of @entity10734 with dilutions of soluble @entity2841 before injection of the mixture over the @entity2841 matrix, prevented ligand binding in a dose dependent manner. In contrast, unrelated soluble cytokines or serum proteins (e.g. @entity1261 , albumin, etc.) were found to exert no inhibition. Complexes formation blockage by pre-incubation of constant amounts of @entity10734 with dilutions of neutralizing anti- @entity10734 antibodies was concentration dependent, further assessing the specificity of the interaction. To investigate the possibility of relating the effect on binding affinity of critical conformational changes at the contact site, experiments of multisite binding were performed, flowing a set of neutralizing monoclonal antibodies reacting to different epitopes on @entity2841 over the @entity2841 matrix, before injecting @entity10734 . The results indicated that antibody interaction with helix D and helix A of @entity2841 markedly inhibited @entity2841 binding to @entity10734 . Comparable results were obtained using the biosensor technology and enzyme-linked immunoassays, in representative experiments performed with the same reagents. These experiments demonstrate that SPR can be successfully used for studying complementary interactions between @entity2841 and its receptor alpha-chains in solution without using labels or secondary tracers and, compared with conventional immunoanalysis methods, significantly saving time.

@entity1 XXXX interaction with the @entity7722 of its receptor studied using surface plasmon resonance.

@entity1 :: ('9606', 'Species') :: ['human'] @entity7722 :: ('2217', 'Gene') :: ['alpha-chain'] @entity1261 :: ('1440', 'Gene') :: ['G-CSF'] @entity6115 :: ('CHEBI:32952', 'Chemical') :: ['amine'] @entity10734 :: ('1438', 'Gene') :: ['GM-Ralpha'] @entity2841 :: ('1437', 'Gene') :: ['granulocyte-macrophage colony-stimulating factor', 'GM-CSF']
@entity2841 :: (1437,Gene) :: ['GM-CSF']
