| General information | |
| ACovPid: | ACoVP100428 |
| Trivial Name: | H |
| Amino Acids Sequence: | HVTTTFAPPPPR |
| Length: | 12 |
| C-Terminal Modification: | None |
| N-Terminal Modification: | None |
| Chemical Modification: | None |
| Peptide Source: | |
| Source Description: | The purified pAPN was used as the target protein and subjected to biopanning using a Ph.D.-12 Phage Display Peptide Library Kit. Sequencing and peptide synthesis:Ten positive phage clones were amplified and precipitated with polyethylene glycol/NaCl. DNA |
| Against Virus: | Transmissible gastroenteritis virus (TGEV) : 11151 |
| Inhibition Value Type: | IC50 |
| Inhibitory Effect: | ~15 |
| Inhibitory Unit: | µM |
| Target Domain Name: | Interaction with TGEV spike glycoprotein domain |
| Assay: | Virus binding blocking assay |
| Assay Description: | Transmissible gastroenteritis virus (TGEV, Purdue 46-MAD strain, a gift from Dr. Luis Enjuanes, Centro Nacional de Biotecnología, CSIC, Madrid, Spain) was used for virus infection assays. To analyze the blockade of the anti-pAPN antibody to TGEV infection, ST cells in 6-well plates were incubated with the antibody achieved in this study at 37 °C for 1 h. After complete washing, TGEV at an MOI (multiplicity of infection) of 10 was used to infect the cells for 1 h. The cells were washed three times with PBS and the inoculums were replaced with 1% (w/v) methylcellulose in EMEM (1 ml/well) followed by incubation for 48–72 h. Unrelated control rabbit sera were included as control. The wells were subjected to virus-plaque reduction assay using crystal violet staining (Li et al., 2009, Ren et al., 2008). |
| Anti-CoV activity in vivo: | |
| Reference: | 21176936 |
| Comment: | Biopanning and enrichment analysis : The purified pAPN was used as the target protein and subjected to biopanning using a Ph.D.-12 Phage Display Peptide Library Kit. The procedure was performed according to the manufacturer's instructions with modification (New England Biolabs, USA). Briefly, the protein of interest at a concentration of 10 µg/well in 0.1 M NaHCO3 (pH 8.6) buffer was coated onto 96-well plates overnight at 4 °C. Then the plates were blocked for 1 h at 4 °C with PBS containing 1% BSA followed by six times washing with TBST (50 mM Tris–HCl, pH 7.5, 150 mM NaCl, 0.1% [vol/vol] Tween 20). The phage library diluted in TBST was added into the plate at a final concentration of 2 × 1011 pfu/ml (100 µl/well) at room temperature for 30 min with gentle rocking. Unbound phage was removed by 10 times washing with TBST. Bound phage was eluted using 100 µl elution buffer (0.2 M glycine–HCl, pH 2.2). The eluates neutralized with 15 µl 1 M Tris–HCl (pH 9.1) were amplified in E. coli ER2738 bacteria. The phages were purified by polyethylene glycol precipitation according to the manufacturer's instructions. Four rounds of panning were performed and the phage concentrations in the elution buffer and after amplification in host cells were |
| 3D structure: | |
| Structure Experiment Verified: | NO |
| Similar Peptides: | |
| Target Domain information | |
| Target Domain Full Name: | Interaction with TGEV spike glycoprotein domain |
| Target Type: | glycoprotein |
| UniprotID [Sequence]: | P15145 [717-813] |
| Target Synonyms: | Alternative name(s): Alanyl aminopeptidase Aminopeptidase M Microsomal aminopeptidase gp130 CD_antigen: CD13 |
| Target Source: | |
| Target Structure: | |