An Engineered Receptor-Binding Domain Improves the Immunogenicity of Multivalent SARS-CoV-2 Vaccines
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (ScreenIT)
Abstract
All available vaccines for coronavirus disease 2019 (COVID-19) express or deliver the full-length SARS-CoV-2 spike (S) protein. We show that this antigen is not optimal, consistent with observations that the vast majority of the neutralizing response to the virus is focused on the S-protein receptor-binding domain (RBD).
Article activity feed
-
-
SciScore for 10.1101/2020.11.18.388934: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: Protein immunizations and sera collection in rats: All animals used in these studies were handled and maintained in accordance with NIH guidelines and approved by Institutional Animal Care and Use Committee (IACUC) of Scripps Research (Protocol 18-025). Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Female Sprague Dawley rats were immunized with incremental increasing doses of antigen over seven days starting at day 0, and boosted with a similar regimen at day 30. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources hACE2 expression was confirmed by SARS1-PV and SARS2-PV … SciScore for 10.1101/2020.11.18.388934: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: Protein immunizations and sera collection in rats: All animals used in these studies were handled and maintained in accordance with NIH guidelines and approved by Institutional Animal Care and Use Committee (IACUC) of Scripps Research (Protocol 18-025). Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Female Sprague Dawley rats were immunized with incremental increasing doses of antigen over seven days starting at day 0, and boosted with a similar regimen at day 30. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources hACE2 expression was confirmed by SARS1-PV and SARS2-PV entry assays and by immunofluorescence staining using mouse monoclonal antibody recognizing c-Myc. c-Mycsuggested: NoneAfter washing, cells were stained with anti-rabbit IgG-Alexa647 antibody for 45 minutes at 4°C, and mean fluorescence intensities were measured for each well by flow cytometry. anti-rabbit IgG-Alexa647suggested: NoneMeasurement of antibody-dependent enhancement: The ability of anti-SARS-CoV-2 RBD immune sera to mediate antibody-dependent enhancement (ADE) was measured using HEK293T cells or HEK293T cells stably expressing human ACE2 (293T-hACE2 cells), transfected using the calcium phosphate transfection method to express the rat ortholog of FcγRI (CD64). anti-SARS-CoV-2suggested: NoneACE2suggested: NoneCD64suggested: NoneExperimental Models: Cell Lines Sentences Resources 293T-hACE2 cells transduced with MLV vectors were selected and maintained with medium containing puromycin (Sigma). 293T-hACE2suggested: NoneProtein Production: Expi293 cells (Thermo-Fisher) were transiently transfected using FectoPRO (Polyplus) with plasmids encoding SARS-CoV2 RBD with a human or rabbit-Fc fusion or a C-terminal C-tag (-EPEA). Expi293suggested: RRID:CVCL_D615)For mouse studies, one hour later, 104 ACE2-239T cells were added and spun at 3000×g for 30 minutes at 4°C, was then returned to 37°C, and media was exchanged 2 hours later with fresh media without mouse sera. ACE2-239Tsuggested: NoneMeasurement of antibody-dependent enhancement: The ability of anti-SARS-CoV-2 RBD immune sera to mediate antibody-dependent enhancement (ADE) was measured using HEK293T cells or HEK293T cells stably expressing human ACE2 (293T-hACE2 cells), transfected using the calcium phosphate transfection method to express the rat ortholog of FcγRI (CD64). HEK293Tsuggested: NoneThe human monocytic cell line K562 (ATCC CCL-243), which endogenously expresses FcγRII, was also used for ADE assays. K562suggested: ATCC Cat# CCL-243, RRID:CVCL_0004)Experimental Models: Organisms/Strains Sentences Resources Female Sprague Dawley rats were immunized with incremental increasing doses of antigen over seven days starting at day 0, and boosted with a similar regimen at day 30. Sprague Dawleysuggested: NoneFemale 8 to 9-week-old BALB/cJ mice were electroporated with 60 μg DNA in each hindquarter for a total dose of 120 μg on day 0 and day 14. BALB/cJsuggested: NoneResults from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
-
