Fig. 1: A novel linear and broadly neutralizing peptide in the S2 protein of SARS-CoV-2. | Cellular & Molecular Immunology

Fig. 1: A novel linear and broadly neutralizing peptide in the S2 protein of SARS-CoV-2.

From: A novel linear and broadly neutralizing peptide in the SARS-CoV-2 S2 protein for universal vaccine development

Fig. 1: A novel linear and broadly neutralizing peptide in the S2 protein of SARS-CoV-2.The alternative text for this image may have been generated using AI.

a ELISA with the indicated peptides as antigens was used to test sera from recovered COVID-19 patients. LOD, limit of detection. b Sequence alignment showing the location of P4 in SARS-CoV-2, SARS-CoV, BatCoV RaTG13 and SARS-CoV-2 variants. c A 3D structure model diagram of the SARS-CoV-2 S protein generated with PyMOL based on PDB 6VSB_1_1_1 [10]. The P4 peptide is shown in red. d and e Neutralizing activity of B10, C10, E2, and G5 mAbs against SARS-CoV-2 S pseudovirus (d) and SARS-CoV S pseudovirus (e). f Neutralizing activity of mAb G5 against the SARS-CoV-2 S pseudovirus B.1.351 South Africa variant. In (d–f), measurements were performed using purified mAbs and SARS-CoV-2 and SARS-CoV S pseudoviruses in hACE2/293T cells, and the results are shown as the percentage of inhibition (mean ± SD). g Neutralizing activity of B10, C10, E2, and G5 mAbs against wild-type SARS-CoV-2. The assay was performed with mouse ascites and wild-type SARS-CoV-2 in Vero E6 cells, and the data are expressed as the log2 number of dilution. h Inhibition of S protein-mediated virus spread by mAbs. i Affinities of B10, C10, E2, and G5 mAbs for the S protein were analyzed by immunoprecipitation assays. 293T cells were transfected with a plasmid carrying the SARS-CoV-2 S gene, and mAbs B10, C10, E2, and G5 were used as the capture antibody to perform immunoprecipitation, followed by western blot analysis using mAb G5. j Identification of the trimeric P4 protein by western blotting. The supernatants and pellets from lysates of BL21 cells transformed with 3×P4 or the pColdI vector were analyzed by western blotting using mAb G5 as the primary antibody. k Neutralization of SARS-CoV-2 pseudovirus by sera from mice immunized with the trimeric P4 protein. The assay was performed with mouse sera and SARS-CoV-2 pseudovirus in hACE2/293T cells. l Inhibition of S-mediated cell-cell fusion by mAbs. m A schematic diagram showing the potential mechanism by which mAbs B10, C10, E2, and G5 neutralize SARS-CoV-2/SARS-CoV. mAbs against 1147SFKEELDKYFKNHTSP1162 inhibit SARS-CoV-2/SARS-CoV viral entry by inhibiting fusion between the virus and cell membrane

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