Recombinant SARS-CoV-2 spike S1-Fc fusion protein induced high levels of neutralizing responses in nonhuman primates

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Abstract

The COVID-19 outbreak has become a global pandemic responsible for over 2,000,000 confirmed cases and over 126,000 deaths worldwide. In this study, we examined the immunogenicity of CHO-expressed recombinant SARS-CoV-2 S1-Fc fusion protein in mice, rabbits, and monkeys as a potential candidate for a COVID-19 vaccine. We demonstrate that the S1-Fc fusion protein is extremely immunogenic, as evidenced by strong antibody titers observed by day 7. Strong virus neutralizing activity was observed on day 14 in rabbits immunized with the S1-Fc fusion protein using a pseudovirus neutralization assay. Most importantly, in less than 20 days and three injections of the S1-Fc fusion protein, two monkeys developed higher virus neutralizing titers than a recovered COVID-19 patient in a live SARS-CoV-2 infection assay. Our data strongly suggests that the CHO-expressed SARS-CoV-2 S1-Fc recombinant protein could be a strong candidate for vaccine development against COVID-19.

Highlights

  • CHO-expressed S1-Fc protein is very immunogenic in various animals and can rapidly induce strong antibody production

  • S1-Fc protein solicits strong neutralizing activities against live virus

  • Stable CHO cell line expressing 50 mg/L of S1-Fc and a 3,000 L Bioreactor can produce 3 million doses of human COVID-19 vaccine every 10 days, making it an accessible and affordable option for worldwide vaccination

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    1. SciScore for 10.1101/2020.04.21.052209: (What is this?)

      Please note, not all rigor criteria are appropriate for all manuscripts.

      Table 1: Rigor

      Institutional Review Board Statementnot detected.
      Randomizationnot detected.
      Blindingnot detected.
      Power Analysisnot detected.
      Sex as a biological variableFemale BALB/c mice were obtained from Vital River Co., China. New Zealand White rabbits were purchased and hosted at Longan Co., China.
      Cell Line Authenticationnot detected.

      Table 2: Resources

      Experimental Models: Cell Lines
      SentencesResources
      Pseudovirus neutralization assay: HEK 293T cells and ACE2-transfected HEK 293T cells (ACE2-293T) were cultured in DMEM (Gibco, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS, Gibco), 50 U/ml penicillin (Gibco) and 50μg/ml streptomycin (Gibco) at 37°C with 5% CO2.
      HEK 293T
      suggested: None
      ACE2-293T cells were seeded in 96-well plates at one day prior to infection.
      ACE2-293T
      suggested: None
      The diluted serums were mixed with SARS-CoV-2 suspension of 100 TCID50 in 96-well plates at a ratio of 1:1, followed by 2 hours incubation at 36.5°C in a 5% CO2 incubator. 1-2 ×104 Vero cells were then added to the serum-virus mixture, and the plates were incubated for 5 days at 36.5°C in a 5% CO2 incubator.
      Vero
      suggested: None
      Experimental Models: Organisms/Strains
      SentencesResources
      Female BALB/c mice were obtained from Vital River Co., China. New Zealand White rabbits were purchased and hosted at Longan Co., China.
      BALB/c
      suggested: RRID:IMSR_ORNL:BALB/cRl)
      Software and Algorithms
      SentencesResources
      The 50% neutralization titer was calculated by probit analysis using the SPSS software.
      SPSS
      suggested: (SPSS, RRID:SCR_002865)

      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 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.
      • 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.

      About SciScore

      SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.