Lactotransferrin (LTF), also known as actoferrin (LF) , is a multifunctional protein of the transferrin family. Lactoferrin belongs to the innate immune system. Apart from its main biological function, namely binding and transport of iron ions, lactoferrin also has antibacterial, antiviral, antiparasitic, catalytic, anti-cancer, and anti-allergic functions and properties. LTF is widely represented in various secretory fluids, such as milk, saliva, tears, and nasal secretions. It also present in secondary granules of PMN and is secreted by some acinar cells. Besides, Protease, Serine 1 (PRSS1) has been identified as an interactor of LTF, thus a binding ELISA assay was conducted to detect the interaction of native bovine LTF and recombinant human PRSS1. Briefly, LTF were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100ul were then transferred to CP-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-LTF 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 50μL stop solution to the wells and read at 450nm immediately. The binding activity of native bovine LTF and recombinant human PRSS1 was shown in Figure 1, the EC50?for this effect is 2.92ug/mL.