rna and human codon-optimized cdna encoding the sars-cov-2 spike protein  (GenScript corporation)

Journal: Nature Communications

Article Title: S-Trimer, a COVID-19 subunit vaccine candidate, induces protective immunity in nonhuman primates

doi: 10.1038/s41467-021-21634-1

Figure Lengend Snippet: a Schematic representations of full-length SARS-CoV-2 Spike (S) protein and the ectodomain of wild-type SARS-CoV-2 S protein-Trimer-Tag fusion protein (S-Trimer). b Schematic 2-D illustration of S-Trimer with homotrimeric Spike protein in the prefusion conformation. c Reducing SDS-PAGE analysis with Coomassie Blue staining of high-level expression of S-Trimer as a secreted protein from CHO cells in a 15L bioreactor Fed-batch serum-free culture over 11 days (10 µL of cleared media were loaded for each sample) along with a purified standard (Std). d S-Trimer is a disulfide bond-linked homotrimer as analyzed by SDS-PAGE with Coomassie Blue staining under non-reducing (-ME) and reducing (+ME) conditions. S-Trimer was shown to be partially cleaved at S1/S2 junction as indicated. e S-Trimer is heavily N -glycosylated. Analysis of S-Trimer before and after deglycosylation with PNGase F (-N) and PNGase F & Endo-O (-O) by SDS-PAGE with Coomassie Blue staining under reducing (+ME) condition. f SEC-HPLC analysis of the purity of S-Trimer with an MW of approximately 700 Kda, and a small fraction of cleaved S1 was shown detached from S-Trimer as indicated. g Determination of the binding affinity between S-Trimer and human ACE2-Fc by ForteBio BioLayer interferometry. All of the above results are representatives of at least three independent experiments.

Article Snippet: To produce the wild-type secreted S-Trimer fusion protein, a cDNA encoding the ectodomain of wild-type SARS-CoV-2 spike (S) protein (amino acid residues 1 to 1211) (GenBank: MN908947.3) was gene-synthesized using Cricetulus griseus (Chinese hamster)-preferred codons by GenScript.

Techniques: SDS Page, Staining, Expressing, Purification, Binding Assay

Journal: NPJ Vaccines

Article Title: Rabies virus-based COVID-19 vaccine CORAVAX™ induces high levels of neutralizing antibodies against SARS-CoV-2

doi: 10.1038/s41541-020-00248-6

Figure Lengend Snippet: a Schematic illustration of CORAVAX, the rabies virus-based SARS-CoV-2 vaccine construct used in this study. A SARS-CoV-2 S1 RABV G chimeric protein cDNA was inserted between the N and P genes of the SAD-B19-derived RABV virus vaccine vector BNSP333. b Immunofluorescence staining of Vero cells at 48 h post-infection labeled for either the RABV G protein (green) or the SARS-CoV-2 S1 protein (red).Scale bar represents 30 μm. c SDS-PAGE analysis of purified virions after sucrose gradient purification. Letters indicate the positions of the RABV L, G, N, P, and M and the chimeric S1-RABV-G proteins. d The panel shows western blotting of purified CORAVAX or control RABV(BNSP333) particles probed with anti-SARS-CoV-2 S rabbit polyclonal antibody or a human monoclonal 4C12 antibody directed against RABV G. All blots derive from the same experiment and were processed in parallel. Full size gels and WB are presented in the .

Article Snippet: Codon-optimized cDNA encoding SARS-CoV-2 Spike protein and RABV glycoprotein were obtained from Genscript.

Techniques: Construct, Derivative Assay, Plasmid Preparation, Immunofluorescence, Staining, Infection, Labeling, SDS Page, Purification, Western Blot

Journal: NPJ Vaccines

Article Title: Rabies virus-based COVID-19 vaccine CORAVAX™ induces high levels of neutralizing antibodies against SARS-CoV-2

doi: 10.1038/s41541-020-00248-6

Figure Lengend Snippet: Balb/C mice were immunized once with live CORAVAX or twice (days 0 and 21) with 10 μg chemically inactivated particles of CORAVAX with or without adjuvant. Serum was collected from each mouse at days 14, 21, 28, 42, and 56 for analysis by a SARS-CoV-2 S-specific ELISA. a Immunization schedule: live CORAVAX used once on day 0; the inactivated vaccines were dosed on days 0 and 28 (figure created with BioRender.com). b SARS CoV-2 S1 IgG responses represented as EC50 titers c SARS CoV-2 RBD IgG responses on day 56 sera d SARS CoV-2 S1 IgG2/1 isotype ratio. Two animals in the inactivated CORAVAX group did not make detectable IgG1 antibodies.

Article Snippet: Codon-optimized cDNA encoding SARS-CoV-2 Spike protein and RABV glycoprotein were obtained from Genscript.

Techniques: Enzyme-linked Immunosorbent Assay