Potent Neutralization Antibodies Induced by a Recombinant Trimeric Spike Protein Vaccine Candidate Containing PIKA Adjuvant for COVID-19
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Abstract
The structures of immunogens that elicit the most potent neutralization antibodies to prevent COVID-19 infection are still under investigation. In this study, we tested the efficacy of a recombinant trimeric Spike protein containing polyI:C (PIKA) adjuvant in mice immunized by a 0–7–14 day schedule. The results showed that a Spike protein-specific antibody was induced at Day 21 with titer of above 50,000 on average, as measured by direct binding. The neutralizing titer was above 1000 on average, as determined by a pseudo-virus using monoclonal antibodies (40592-MM57 and 40591-MM43) with IC50 at 1 μg/mL as standards. The protein/peptide array-identified receptor-binding domain (RBD) was considered as immunodominant. No linear epitopes were found in the RBD, although several linear epitopes were found in the C-terminal domain right after the RBD and heptad repeat regions. Our study supports the efficacy of a recombinant trimeric Spike protein vaccine candidate for COVID-19 that is safe and ready for storage and distribution in developing countries.
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SciScore for 10.1101/2021.02.17.431647: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: Vaccination in mice: Animal experiments were approved by the institutional board of Tongji University School of Medicine. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources The arrays were washed with PBST and bound antibodies were detected by incubating with Cy3-conjugated goat anti-mouse IgG (Jackson ImmunoResearch, PA, USA), which were diluted for 1: 1,000 in PBST. anti-mouse IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources The plasmid was transfected to 293T cells and the recombinant S protein … SciScore for 10.1101/2021.02.17.431647: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: Vaccination in mice: Animal experiments were approved by the institutional board of Tongji University School of Medicine. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources The arrays were washed with PBST and bound antibodies were detected by incubating with Cy3-conjugated goat anti-mouse IgG (Jackson ImmunoResearch, PA, USA), which were diluted for 1: 1,000 in PBST. anti-mouse IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources The plasmid was transfected to 293T cells and the recombinant S protein trimers were purified by Ni-NTA (nickel-nitrilotriacetic acid) chromatography (QIAGEN, Germany), followed by size exclusion to further purify the trimers. 293Tsuggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)The plasmids of 9μg pHAGE-luciferase-GFP、 psPAX2 and Zhou-COVID-19-Spike were co-transfected into HEK293T cells by using 1 μg/mL polyetherimide (Polysciences, Warrington, PA) in DMEM medium containing 10% FCS. HEK293Tsuggested: NoneThe mixtures of pseudo-viruses and antibodies were added to 293T/ACE2 cells. 293T/ACE2suggested: RRID:CVCL_YZ65)Experimental Models: Organisms/Strains Sentences Resources C57BL/6 mice at 6 to 8 weeks old were immunized by intramuscular injection of PIKA-S-trimer vaccine or PIKA-S-monomer vaccine. C57BL/6suggested: NoneSoftware and Algorithms Sentences Resources The absorbance was measured at 450 nm and the antibody titer was calculated with GraphPad Prism 7.0 (San Diego, CA). GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)The 50% neutralization dose was calculated using GraphPad Prism 7.0 (San Diego, CA). GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)Results 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.
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