Mice Immunized with the Vaccine Candidate HexaPro Spike Produce Neutralizing Antibodies against SARS-CoV-2
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (ScreenIT)
Abstract
Updated and revised versions of COVID-19 vaccines are vital due to genetic variations of the SARS-CoV-2 spike antigen. Furthermore, vaccines that are safe, cost-effective, and logistic-friendly are critically needed for global equity, especially for middle- to low-income countries. Recombinant protein-based subunit vaccines against SARS-CoV-2 have been reported using the receptor-binding domain (RBD) and the prefusion spike trimers (S-2P). Recently, a new version of prefusion spike trimers, named HexaPro, has been shown to possess two RBD in the “up” conformation, due to its physical property, as opposed to just one exposed RBD found in S-2P. Importantly, this HexaPro spike antigen is more stable than S-2P, raising its feasibility for global logistics and supply chain. Here, we report that the spike protein HexaPro offers a promising candidate for the SARS-CoV-2 vaccine. Mice immunized by the recombinant HexaPro adjuvanted with aluminum hydroxide using a prime-boost regimen produced high-titer neutralizing antibodies for up to 56 days after initial immunization against live SARS-CoV-2 infection. Also, the level of neutralization activity is comparable to that of convalescence sera. Our results indicate that the HexaPro subunit vaccine confers neutralization activity in sera collected from mice receiving the prime-boost regimen.
Article activity feed
-
-
SciScore for 10.1101/2021.02.27.433054: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Mouse immunization: Female C57BL/6 mice (7-9 weeks old, n = 3 per group) were ordered from Nomura Siam International. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources A goat anti-rabbit secondary antibody (IgG) conjugated with Alexa Fluor 594 (Invitrogen, Cat#A-11037) or a goat anti-mouse secondary antibody (IgG) conjugated with Alexa Fluor 488 (Invitrogen, Cat#A-11029) was used for visualization under a fluorescence microscope. anti-rabbitsuggested: (Thermo Fisher Scientific Cat# A-11037, RRID:AB_2534095)anti-mousesug…SciScore for 10.1101/2021.02.27.433054: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Mouse immunization: Female C57BL/6 mice (7-9 weeks old, n = 3 per group) were ordered from Nomura Siam International. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources A goat anti-rabbit secondary antibody (IgG) conjugated with Alexa Fluor 594 (Invitrogen, Cat#A-11037) or a goat anti-mouse secondary antibody (IgG) conjugated with Alexa Fluor 488 (Invitrogen, Cat#A-11029) was used for visualization under a fluorescence microscope. anti-rabbitsuggested: (Thermo Fisher Scientific Cat# A-11037, RRID:AB_2534095)anti-mousesuggested: (Molecular Probes Cat# A-11029, RRID:AB_138404)For convalescent serum staining, cells were incubated with heat-inactivated serum and visualized with a goat anti-human secondary antibody (IgG) conjugated with FITC (Abcam, Cat#ab97224) anti-human secondary antibody ( IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources HEK293T cells were transiently transfected with the HexaPro plasmid by calcium phosphate transfection. HEK293Tsuggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)Immunofluorescence staining: HeLa cells were transiently transfected with the plasmid encoding HexaPro using lipofectamine 3000 (Invitrogen, Cat#L3000008). HeLasuggested: NoneAll the experiments will be used live SARS-CoV–2 viruses at passage 3 or 4 with the Vero E6 cells at the maximum passages of 20. Vero E6suggested: NoneExperimental Models: Organisms/Strains Sentences Resources Mouse immunization: Female C57BL/6 mice (7-9 weeks old, n = 3 per group) were ordered from Nomura Siam International. C57BL/6suggested: NoneSoftware and Algorithms Sentences Resources Supernatant were concentrated with Amicon® Ultra–15 Ultrace–30K centrifugal filter unit (MERCK). Amicon®suggested: 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: We found the following clinical trial numbers in your paper:
Identifier Status Title NCT04522089 Recruiting A Study to Evaluate the Safety and Immunogenicity of COVID-1… NCT04527575 Active, not recruiting Study of the Safety, Reactogenicity and Immunogenicity of "E… NCT04683484 Recruiting A Clinical Trial to Assess the Safety and Immunogenicity of … NCT04742738 Recruiting Safety and Immunogenicity Study of SARS-CoV-2 Nanoparticle V… NCT0464148 Trial number did not resolve on clinicaltrials.gov. Is the number correct? NA 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.
-
