40591 v08h  (Sino Biological)

Journal: bioRxiv

Article Title: Identification of four linear B-cell epitopes on the SARS-CoV-2 spike protein able to elicit neutralizing antibodies

doi: 10.1101/2020.12.13.422550

Figure Lengend Snippet: The predicted linear B-cell epitopes in the Spike protein of SARS-CoV-2. a , The number of linear B-cell epitopes shared among the distinct methods and literature mining. The pink, green and light blue represent epitopes with antigenicity scores > 0.9, 0.4 and 0.9, and

Article Snippet: SARS-CoV-2 S1-specific IgG assay in horse, mouse, monkey and human (Indirect ELISA)96-well polystyrene microplates (Oriental Ocean Global Health, China) were coated with 2 μg/mL (50μL/well) SARS-CoV-2 Spike Protein (S1 Subunit, His tag) (Sino Biological, China, cat no:40591-V08H) in carbonate bicarbonate buffer pH9.6 and the plates were incubated at 4°C overnight.

Techniques:

Journal: bioRxiv

Article Title: Identification of four linear B-cell epitopes on the SARS-CoV-2 spike protein able to elicit neutralizing antibodies

doi: 10.1101/2020.12.13.422550

Figure Lengend Snippet: Measurements of the selected Linear B cell epitope binding to antibody and neutralization efficiency of selected epitopes against SARS-CoV-2. a-d, The binding affinity assessed by ELISA between linear B-cell epitopes and serum antibodies from immunized horse with S1-based vaccines (a), immunized mouse with RBD-based vaccines (b), immunized monkey with RBD-based vaccines (c), and a patient recovering from COVID-19 (d). e, The binding affinity assessed by ELISA between the linear B-cell epitopes and serum antibodies from immunized mice with corresponding epitopes of ‘YNSASFSTFKCYGVSPTKLNDLCFT’, ‘GDEVRQIAPGQTGKIADYNYKLP’, ‘YQPYRVVVLSFELLH’, and ‘CVNFNFNGL’. f, Neutralization assay against SARS-CoV-2 pseudovirus in ‘YNSASFSTFKCYGVSPTKLNDLCFT’, ‘GDEVRQIAPGQTGKIADYNYKLP’, ‘YQPYRVVVLSFELLH’, and ‘CVNFNFNGL’. y-axis is the value of EC 50 . g, Neutralization assay against SARS-CoV-2 live virus in ‘YNSASFSTFKCYGVSPTKLNDLCFT’, ‘GDEVRQIAPGQTGKIADYNYKLP’, ‘YQPYRVVVLSFELLH’, and ‘CVNFNFNGL’. y-axis is the value of NT 50 .

Article Snippet: SARS-CoV-2 S1-specific IgG assay in horse, mouse, monkey and human (Indirect ELISA)96-well polystyrene microplates (Oriental Ocean Global Health, China) were coated with 2 μg/mL (50μL/well) SARS-CoV-2 Spike Protein (S1 Subunit, His tag) (Sino Biological, China, cat no:40591-V08H) in carbonate bicarbonate buffer pH9.6 and the plates were incubated at 4°C overnight.

Techniques: Binding Assay, Neutralization, Enzyme-linked Immunosorbent Assay, Mouse Assay

Journal: bioRxiv

Article Title: Identification of four linear B-cell epitopes on the SARS-CoV-2 spike protein able to elicit neutralizing antibodies

doi: 10.1101/2020.12.13.422550

Figure Lengend Snippet: The characteristics of the 18 selected linear B cell epitopes. a , The sequences of 18 selected linear B-cell epitopes. The bold is the mutated site in less than ten of 118,694 virus strains; The red is the predicted discontinuous residues. The bars on the right side are the Wilcoxon test p value for the comparisons of IgG or IgA antibody enrichment scores associated with each linear B-cell epitope between COVID-19 patients and negative controls. b , The digesting enzymes profile of the epitope sequence. Red indicated not digest, blue indicated digest. c-d , The localization of the 18 selected epitopes mapped on SARS-CoV-2 S (PDB: 6VSB) protein (c) and ACE-RBD complex (d). e-f , The localizations of B cell discontinuous epitopes on SARS-CoV-2 S (PDB: 6VSB) protein (e) and ACE-RBD complex (f). The spike protein is grey, the RBD region is wheat color, the selected epitopes are green, the mutation sites are red, the human ACE domain is blue, and the discontinuous B-cell epitopes are purple.

Article Snippet: SARS-CoV-2 S1-specific IgG assay in horse, mouse, monkey and human (Indirect ELISA)96-well polystyrene microplates (Oriental Ocean Global Health, China) were coated with 2 μg/mL (50μL/well) SARS-CoV-2 Spike Protein (S1 Subunit, His tag) (Sino Biological, China, cat no:40591-V08H) in carbonate bicarbonate buffer pH9.6 and the plates were incubated at 4°C overnight.

Techniques: Sequencing, Mutagenesis

Journal: Animal Models and Experimental Medicine

Article Title: Age‐related rhesus macaque models of COVID‐19, et al. Age‐related rhesus macaque models of COVID‐19

doi: 10.1002/ame2.12108

Figure Lengend Snippet: Viral load of the SARS‐CoV‐2‐infected rhesus macaque model. A. Average viral loads of swabs from the younger group (YG, n = 3, red line) monkeys. B. Average viral load of swabs from the elder group (EG, n = 2, blue line) monkeys. Viral loads of nasal, throat, and anal swab specimens collected from the inoculated macaques on 0, 3, 5, 7, 9, 11, and 14 dpi. C. Viral loads in varies lobe of lung tissue from YG and EG monkeys at day 7 post‐inoculation. RNA was extracted and viral load was determined by qRT‐PCR. All data are presented as mean ± SEM

Article Snippet: The 96‐well plates coated with 0.1 μg Spike protein from SARS‐CoV‐2 (Sino Biological; Product code:40591‐V08H) at 4℃ overnight were blocked by 2% BSA/PBST at room temperature for 1 hour.

Techniques: Infection, Quantitative RT-PCR

Journal: Animal Models and Experimental Medicine

Article Title: Age‐related rhesus macaque models of COVID‐19, et al. Age‐related rhesus macaque models of COVID‐19

doi: 10.1002/ame2.12108

Figure Lengend Snippet: Hematological analysis in rhesus macaques inoculated with SARS‐CoV‐2. A. The counts of white blood cells (WBC) were analysed. B. The percentage and counts of monocytes were determined. C. The percentage and counts of lymphocytes were detected. D. The percentage and counts of CD3 + CD8 + T cells, CD3 + CD4 + T cells were shown. YG (red line) and EG (blue line) were indicated in the upper right corner of each panel. All data are presented as mean ± SEM

Article Snippet: The 96‐well plates coated with 0.1 μg Spike protein from SARS‐CoV‐2 (Sino Biological; Product code:40591‐V08H) at 4℃ overnight were blocked by 2% BSA/PBST at room temperature for 1 hour.

Techniques:

Journal: Animal Models and Experimental Medicine

Article Title: Age‐related rhesus macaque models of COVID‐19, et al. Age‐related rhesus macaque models of COVID‐19

doi: 10.1002/ame2.12108

Figure Lengend Snippet: The comparison of lesions in the lung between younger group (YG) and elder group (EG) by radiographic alterations, histopathological and immunohistochemical (IHC) observation of the SARS‐CoV‐2‐inoculated‐rhesus macaque. A. Anterior‐posterior thoracic X‐rays from of rhesus macaque imaged prior to SARS‐CoV‐2 inoculation (day 0) and on 7 dpi of YG and 5 dpi of EG. Areas of interstitial infiltration, indicative of pneumonia, are highlighted (red circle). Positional indicators are included (R = right). B. Histopathological changes in lungs from YG and EG. Lung tissue was collected and stained with hematoxylin and eosin. Black scale bar = 40 µm. IHC staining demonstrated that SARS‐CoV‐2 antigens were mainly in the epithelial cells and macrophages. SARS‐CoV‐2 antigens were indicated by red arrows. Red scale bar = 50 µm

Article Snippet: The 96‐well plates coated with 0.1 μg Spike protein from SARS‐CoV‐2 (Sino Biological; Product code:40591‐V08H) at 4℃ overnight were blocked by 2% BSA/PBST at room temperature for 1 hour.

Techniques: Immunohistochemistry, Staining

Journal: bioRxiv

Article Title: Oral delivery of SARS-CoV-2 DNA vaccines using attenuated Salmonella typhimurium as a carrier in rat

doi: 10.1101/2020.07.23.217174

Figure Lengend Snippet: pcDNA3.1(+)-CMV-SARS-CoV-2-S-GFP plasmid map.

Article Snippet: Detection of SARS-CoV-2-S protein expression in vitroFor the western blot detection of SARS-CoV-2-S protein, the cells were washed twice with ice-cold PBS and harvested from the dishes with a cell scraper by adding a WIP lysis buffer.

Techniques: Plasmid Preparation

Journal: bioRxiv

Article Title: Oral delivery of SARS-CoV-2 DNA vaccines using attenuated Salmonella typhimurium as a carrier in rat

doi: 10.1101/2020.07.23.217174

Figure Lengend Snippet: Micrographs of 293T cells transfected with pSARS-CoV-2-S (X 100). (A1, A2) 293T cells transfected with pSARS-CoV-2-S (pcDNA3.1(+)-CMV-SARS-CoV-2-S-GFP) at 48 hours after transfection. A1 fluorescence micrograph with GFP expression in cells and light micrograph with the same visual field as A2. (B) 293T cells transfected with pSARS-CoV-2-S at 48 hours after transfection. The SARS-CoV-2-S protein showed about 141 kDa.

Article Snippet: Detection of SARS-CoV-2-S protein expression in vitroFor the western blot detection of SARS-CoV-2-S protein, the cells were washed twice with ice-cold PBS and harvested from the dishes with a cell scraper by adding a WIP lysis buffer.

Techniques: Transfection, Fluorescence, Expressing

Journal: bioRxiv

Article Title: Oral delivery of SARS-CoV-2 DNA vaccines using attenuated Salmonella typhimurium as a carrier in rat

doi: 10.1101/2020.07.23.217174

Figure Lengend Snippet: Humoral responses to SARS-CoV-2-S protein antigen in the rat after immunizationon day 0, day 14, and day 28 with Salmonella carrying the control vector or pSARS-CoV-2-S (as described in the methods). (A) After immunization with the control vector, test SARS-CoV-2-S protein antigen binding of IgG in serial serum dilutions from a rat at day (0, 14, 28). Data shown represent test mean OD450 nm values (mean±SD) for each of 9 rats, or (B) After immunization with the pSARS-CoV-2-S vector, SARS-CoV-2-S protein antigen binding of IgG in serial serum dilutions from a rat at day (0, 14, 28). Data shown represent mean OD450 nm values (3 times measurement, mean±SD) for each of 9 rats.

Article Snippet: Detection of SARS-CoV-2-S protein expression in vitroFor the western blot detection of SARS-CoV-2-S protein, the cells were washed twice with ice-cold PBS and harvested from the dishes with a cell scraper by adding a WIP lysis buffer.

Techniques: Plasmid Preparation, Binding Assay

Journal: bioRxiv

Article Title: D614G mutation of SARS-CoV-2 spike protein enhances viral infectivity

doi: 10.1101/2020.06.20.161323

Figure Lengend Snippet: The S-G614 protein pseudotyped virus showed increased infectivity. a HEK 293T and 293T-ACE2 (human angiotensin-converting enzyme 2) cells were infected with lentiviruses pseudotyped with VSV-G and SARS-CoV-2 S protein variants. Virus titers were quantified by RT-qPCR and adjusted to 3.8 × 10 4 copies in 50 μL to normalize input virus doses. The relative luminescence units (RLU) detected 72 h post-infection (hpi). b Inhibition of pseudoviral entry by ACE2-Ig. Pseudoviruses were pre-incubated with ACE2-Ig and added to 293T-ACE2 cells, then RLU was measured at 72 hpi. c Viral entry efficiency meditated by S variants. The RLU was measured at 24-72 hpi. d-e D614G mutation facilitates elastase-2 induced pseudoviral entry. 293T-ACE2 cells were treated with elastase for 5 min and then infected with pseudotyped viruses containing the S-D614 or S-G614 mutant in the presence of various concentrations of sivelestat sodium. RLU was measured at 72 hpi. n = 3, ±SD. * P

Article Snippet: Antibodies and inhibitorsThe anti-RBD monoclonal antibody against the SARS-CoV-2 S protein was kindly provided by Prof. Aishun Jin from Chongqing Medical University.

Techniques: Infection, Quantitative RT-PCR, Inhibition, Incubation, Mutagenesis