| General information | |
| ACovPid: | ACoVP100516 |
| Trivial Name: | P9 |
| Amino Acids Sequence: | CANLLLQYGSFCTQLNRALSGIA |
| Length: | 23 |
| C-Terminal Modification: | None |
| N-Terminal Modification: | None |
| Chemical Modification: | None |
| Peptide Source: | Severe acute respiratory syndrome coronavirus (SARS-CoV) : 694009 |
| Source Description: | From S1 subunit of spike (S) protein of SARS-CoV |
| Against Virus: | Severe acute respiratory syndrome coronavirus (SARS-CoV) : 694009 |
| Inhibition Value Type: | IC50 |
| Inhibitory Effect: | |
| Inhibitory Unit: | |
| Target Domain Name: | |
| Assay: | Syncytia inhibition assay |
| Assay Description: | Syncytia formation was used to mimic the SARS‐CoV infection procedure. In our study, we modified this assay by introducing luciferase assay in the detecting system. The cell fusion‐dependent reporter gene (luciferase) activation assay was adapted to our studies of spike protein‐mediated membrane fusion. In brief, effector cells were generated by cotransfection of HEK293 T cell monolayers (106 cells/60 mm dish) with plasmids pCDNA3.1‐ACE2 and pUHD 15‐1(SV40), while target cells (containing the luciferase reporter gene) were generated by cotransfection of HEK293 T cell with pCDNA3.1‐spike‐Ig and pUHD 10‐3. Twenty‐four hours after transfection, effector cells and target cells were trypsinized, resuspended in DMEM–10% FBS, then two kinds of cell suspensions were mixed (1:1 cell number) and repelleted. All the antisera and peptides (20 µl of each antiserum and 125 µM of each peptide) were added into the cell suspension mixture to investigate their inhibitory capacity. Antiserum or peptide was incubated with cell suspensions at 23 °C for 1 h, and then re‐plated into a 48‐well culture plate at 37 °C for 24 h. Finally, media were removed and cells were lysed with 100 µl Passive Lysis Buffer (Promega). Luciferase intensity was measured in a luminometer TD‐20/20. The reactivity of the peptides to the SARS patients’ sera was also measured by ELISA. The 96‐well plates were coated with the BSA–peptide conjugates(5 µg/ml, 50 µl/well) in coating buffer (pH 9.4) at 4 °C overnight. Afterwards the wells were washed with PBS, and were blocked with BSA at 37 °C for 2 h and then the sera or the purified IgG from SARS patients’ sera was added as the first Ab. The sera dilution was 1:10 and the purified IgG dilution was 1:1000. Plates were stored at 37 °C for 30 min, then washed again with PBS. After that, HRP‐coupled goat anti‐human IgG (Bio‐rad) was added, and OD values were measured with a microplate autoreader (Biotek) at 450 nm. |
| Anti-CoV activity in vivo: | |
| Reference: | 15811330 |
| Comment: | |
| 3D structure: | |
| Structure Experiment Verified: | NO |
| Similar Peptides: | ACoVP100202 ACoVP37.7 |