Hepatocyte growth factor (HGF) is a paracrine cellular growth, motility and morphogenic factor. It is secreted by mesenchymal cells and targets and acts primarily upon epithelial cells and endothelial cells, but also acts on haemopoietic progenitor cells and T cells. It has been shown to have a major role in embryonic organ development, specifically in myogenesis, in adult organ regeneration, and in wound healing. Besides, Matrix Metalloproteinase 9 (MMP9) has been identified as an interactor of HGF, thus a binding ELISA assay was conducted to detect the interaction of recombinant human HGF and recombinant mouse MMP9. Briefly, HGF were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 ul were then transferred to MMP9 -coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti- HGF pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution , wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450 nm immediately. The binding activity of recombinant human HGF and recombinant mouse MMP9 was shown in Figure 1, the EC50 for this effect is 0.35 ug/mL.