Figure. The binding activity of IL28B with IL10Rb.
Interleukin 28B (IL28B) is a cytokine distantly related to type I interferons and the IL-10 family. It has function on antiviral, antitumour and immunomodulatory activities. IL28B also plays a critical role in the antiviral host defense, predominantly in the epithelial tissues and acts as a ligand for the heterodimeric class II cytokine receptor composed of IL10RB and IFNLR1, and receptor engagement leads to the activation of the JAK/STAT signaling pathway resulting in the expression of IFN-stimulated genes (ISG), which mediate the antiviral state. Besides, Interleukin 10 Receptor Beta (IL10Rb) has been identified as an interactor of IL28B, thus a binding ELISA assay was conducted to detect the interaction of recombinant human IL28B and recombinant human IL10Rb. Briefly, IL28B were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to IL10Rb-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-IL28B 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 IL28B and IL10Rb was shown in Figure 1, and this effect was in a dose dependent manner.