武漢云克隆科技股份有限公司

Angiotensin-converting enzyme 2 (ACE2), as a transmembrane protein, serves as the main entry point into cells for some coronaviruses, including HCoV-NL63, SARS-CoV , and SARS-CoV-2 . More specifically, the binding of the spike S1 protein of SARS-CoV and SARS-CoV-2 to the enzymatic domain of ACE2 on the surface of cells results in endocytosis and translocation of both the virusand the enzyme into endosomes located within cells. Besides, recent studies show that spike (S) proteins of 2019-nCoV and SARS-CoV may use the same host cell receptor called angiotensin-converting enzyme 2 (ACE2) for entering into host cells, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse ACE2 and recombinant Spike glycoprotein(S1). Briefly, ACE2 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to S1-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-ACE2 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 ACE2 and S1 was shown in Figure 1, and this effect was in a dose dependent manner.