NRG1 (neuregulin-1) is a membrane glycoprotein that belongs to the neuregulin family and act on the EGFR family of receptors. It mediates cell-cell signaling and plays a critical role in the growth and development of multiple organ systems. It is reported that by binding to HER3 receptor, NRG1 mediates downstream signaling pathways, leading to multiple effects including growth, proliferation, decreased apoptosis, cellular migration and angiogenesis. Besides NRG1 is direct ligand for ERBB3 and ERBB4 tyrosine kinase receptors and it concomitantly recruits ERBB1 and ERBB2 coreceptors, resulting in ligand-stimulated tyrosine phosphorylation and activation of the ERBB receptors. A functional binding ELISA assay was conducted to detect the interaction of recombinant human NRG1 and recombinant human ERBB2. Briefly, NRG1 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to ERBB2-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-NRG1 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 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 450/630 nm immediately. The binding activity of recombinant human NRG1 and recombinant human ERBB2 was shown in Figure 1, the EC50?for this effect is 0.09 ug/mL.