Protein-based RBD-C-tag COVID-19 Vaccination Candidate Elicits Protection Activity against SARS-COV-2 Variant Infection
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Abstract
The identification of a vaccination candidate against COVID-19 providing protecting activity against emerging SARS-COV-2 variants remains challenging. Here, we report protection activity against a spectrum of SARS-COV-2 and variants by immunization with protein-based recombinant RBD-C-tag administered with aluminum-phosphate adjuvant intramuscularly. Immunization of C57BL/6 mice with RBD-C-tag resulted in the in vivo production of IgG antibodies recognizing the immune-critical spike protein of the SARS-COV-2 virus as well as the SARS-COV-2 variants alpha (“United Kingdom”), beta (“South Africa”), gamma (“Brazil/Japan”), and delta (“India”) as well as wt -spike protein. RBD-C-tag immunization led to a desired Th1 polarization of CD4 T cells producing IFNγ. Importantly, RBD-C-tag immunization educated IgG production delivers antibodies that exert neutralizing activity against the highly transmissible SARS-COV-2 virus strains “Washington”, “South Africa” (beta), and “India” (delta) as determined by conservative infection protection experiments in vitro. Hence, the protein-based recombinant RBD-C-tag is considered a promising vaccination candidate against COVID-19 and a broad range of emerging SARS-COV-2 virus variants.
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SciScore for 10.1101/2021.08.17.456704: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: Study approval: Mouse care and experimental procedures with mice were performed under pathogen-free conditions in accordance with established institutional guidance and approved Animal Care and Use Protocols (ACUP) from the Research Animal Care Committee at Sorrento Therapeutics Inc. Sex as a biological variable Mice and cell culture: For intramuscular antigen challenge, C57BL/6 female mice 5-6 weeks old were injected with either 100 μg RBD-C-tag (300 μg adjuvant) or 50 μg RBD-C-tag (150 μg adjuvant) intramuscularly. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Detection of RBD … SciScore for 10.1101/2021.08.17.456704: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: Study approval: Mouse care and experimental procedures with mice were performed under pathogen-free conditions in accordance with established institutional guidance and approved Animal Care and Use Protocols (ACUP) from the Research Animal Care Committee at Sorrento Therapeutics Inc. Sex as a biological variable Mice and cell culture: For intramuscular antigen challenge, C57BL/6 female mice 5-6 weeks old were injected with either 100 μg RBD-C-tag (300 μg adjuvant) or 50 μg RBD-C-tag (150 μg adjuvant) intramuscularly. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Detection of RBD and Spike binding serum IgG antibodies by ELISA: A direct binding ELISA format was used to detect the anti-SARS-COV-2 RBD or Spike IgG antibody in mouse serum samples. Spike binding serum IgGsuggested: Noneanti-SARS-COV-2 RBD or Spike IgGsuggested: Nonedonkey anti-mouse IgG conjugated to AlexaFluor 647 (Invitrogen #A-31571) or IL-10 APC antibodies ( eBioscience, # 17-7101-81). donkey anti-mouse IgGsuggested: (Molecular Probes Cat# A-31571, RRID:AB_162542)anti-mouse IgGsuggested: (Molecular Probes Cat# A-31571, RRID:AB_162542)IL-10 APCsuggested: (Thermo Fisher Scientific Cat# 17-7101-81, RRID:AB_469501)Experimental Models: Cell Lines Sentences Resources Human parental HEK293 cells (ATCC, CRL-1573) were cultured in DMEM supplemented with 10% FBS and antibiotics/antimycotics (Gibco). HEK293suggested: NoneSimian VeroE6 cells were plated at 18×103 cells/well in a flat bottom 96-well plate in a volume of 200 μl/well. VeroE6suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)Experimental Models: Organisms/Strains Sentences Resources Longitudinal near infrared in vivo imaging of lymphatics: IRDye800-RBD-C-tag was injected intramuscularly into the biceps femoris of C57BL/6 mice. C57BL/6suggested: 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 found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
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.
- No funding statement was detected.
- No protocol registration statement was detected.
Results from scite Reference Check: We found no unreliable references.
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