Inhibition Of Hotspot Peptide Ligand(s) Binding To Recombinant Human Soluble TNFR2 By Anti-TNFR2 Antibody, mAb226. HotSpot Peptide Ligand phage clones were added to 96 well plates with immobilized TNFR2 at a concentration which allowed 50% occupancy of receptor binding sites. Anti-TNFR2 antibody, mAb226, was added and serially diluted for competition. To visualize receptor bound HSPL-phage the reaction was probed with HRP labeled anti-M13 antibody. To demonstrate mAb226 interacted with the TNF binding pocket, TNF-alpha was added in a manner as described for the phage clones and TNF binding was determined using an anti-TNF antibody followed by HRP-anti-mouse IgG antibody. Data is presented as percent inhibition and were fit to a four-parameter logistic equation using Xlfit™, an add-in for Microsoft Excel.