mRNA-based SARS-CoV-2 vaccine candidate CVnCoV induces high levels of virus-neutralising antibodies and mediates protection in rodents

Article activity feed

1. Version published to 10.1038/s41541-021-00311-w

   Apr 16, 2021
2. 

   ScreenIT

   Mar 1, 2021

   SciScore for 10.1101/2020.10.23.351775: (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	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Specific proteins were detected using rabbit anti-SARS Spike c-terminal (Abcam, Cat. ab252690) and mouse anti tubulin (Abcam, Cat. ab7291), followed by goat anti-rabbit IgG IRDye® 800CW (Li-Cor, Cat. 926-32211) and goat anti-mouse IgG IRDye® 680RD (Li-Cor, Cat. 926-68070), respectively.	anti tubulin suggested: None
   Specific S protein expression was assessed via staining with Human anti SARS CoV S antibody (CR3022)	anti SARS CoV S suggested: None CR3022 suggested:…

   SciScore for 10.1101/2020.10.23.351775: (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	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Specific proteins were detected using rabbit anti-SARS Spike c-terminal (Abcam, Cat. ab252690) and mouse anti tubulin (Abcam, Cat. ab7291), followed by goat anti-rabbit IgG IRDye® 800CW (Li-Cor, Cat. 926-32211) and goat anti-mouse IgG IRDye® 680RD (Li-Cor, Cat. 926-68070), respectively.	anti tubulin suggested: None
   Specific S protein expression was assessed via staining with Human anti SARS CoV S antibody (CR3022)	anti SARS CoV S suggested: None CR3022 suggested: None
   MRO-1214LC) followed by goat anti-human IgG F(ab’)2 fragment PE antibody (Immuno Research, Cat. 109-116-097) in a BD FACS Canto II cell analyzer and the FlowJo 10 software.	anti-human IgG suggested: (Jackson ImmunoResearch Labs Cat# 109-116-097, RRID:AB_2337677)
   Antibody analysis: Blood samples were taken via retro-orbital bleeding and SARS-CoV-2 S-specific IgG1 and IgG2a antibodies (mice) or total IgG (hamster) antibodies were detected via ELISA.	SARS-CoV-2 S-specific IgG1 suggested: None S-specific suggested: None IgG2a suggested: None total IgG suggested: None
   For detection, mouse sera were incubated with biotin rat anti-mouse IgG1 (BD Pharmingen, Cat. 550331) or biotin rat anti-mouse IgG2a (BD Pharmingen, Cat. 550332), hamster sera with biotin goat anti-hamster (Syrian) IgG antibody (BioLegend, Cat: 405601) followed by incubation with HRP-Streptavidin (BD, Cat: 554066).	anti-mouse IgG1 suggested: None anti-mouse IgG2a suggested: None anti-hamster ( Syrian ) IgG suggested: None
   The cells were then incubated with 200 ng/well of R-PE labelled rabbit anti-SARS-CoV-2 spike neutralizing monoclonal antibody (Sino Biological, Cat: 40592-R001) at 4°C for 2 h.	anti-SARS-CoV-2 suggested: None
   Antibody labelling with R-PE using the Zenon™ labelling kit for rabbit IgG (Invitrogen, Cat: Z25355) was performed according to the manufacturer’s instructions.	rabbit IgG suggested: None
   The following antibodies were used for flow cytometry analysis: anti-Thy1.2 FITC (clone 53-2.1; Biolegend, Cat.14304)	anti-Thy1.2 FITC suggested: (Abcam Cat# ab24905, RRID:AB_448475)
   Experimental Models: Cell Lines
   Sentences	Resources
   For detection of mRNA expression in cell culture, HeLa cells were seeded in 6-well plates at a density of 400.000 cells/well. 24h later, cells were transfected with 2μg of mRNA per well via Lipofection.	HeLa suggested: None
   Quadruplicate, 10-fold serial dilutions were transferred to 96 well plates with Vero E6 cell culture monolayers and incubated for one hour at 37°C.	Vero E6 suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Mice (BALB/c, 8-12 weeks of age) were obtained from Janvier Laboratories (Le Genest-Saint-Isle, France).	BALB/c suggested: None
   Software and Algorithms
   Sentences	Resources
   MRO-1214LC) followed by goat anti-human IgG F(ab’)2 fragment PE antibody (Immuno Research, Cat. 109-116-097) in a BD FACS Canto II cell analyzer and the FlowJo 10 software.	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   Antibody labelling with R-PE using the Zenon™ labelling kit for rabbit IgG (Invitrogen, Cat: Z25355) was performed according to the manufacturer’s instructions.	Zenon™ suggested: None

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   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).

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   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.

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   Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

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   Results from JetFighter: We did not find any issues relating to colormaps.

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   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.

   ---

   Read the original source
3. 

   ScreenIT

   Oct 26, 2020

   SciScore for 10.1101/2020.10.23.351775: (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

   Female Balb/c mice (n=8/group) were vaccinated IM on day 0, day 7, day 14 or day 21 with 2 µg of CVnCoV.

   Cell Line Authentication

   not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Specific proteins were detected using rabbit anti-SARS Spike c-terminal (Abcam, Cat. ab252690) and mouse anti tubulin (Abcam, Cat. ab7291), followed by goat anti-rabbit IgG IRDye® 800CW (Li-Cor, Cat. 926-32211) and goat anti-mouse IgG IRDye® 680RD (Li-Cor, Cat. 926-68070), respectively.	anti tubulin …

   SciScore for 10.1101/2020.10.23.351775: (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

   Female Balb/c mice (n=8/group) were vaccinated IM on day 0, day 7, day 14 or day 21 with 2 µg of CVnCoV.

   Cell Line Authentication

   not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Specific proteins were detected using rabbit anti-SARS Spike c-terminal (Abcam, Cat. ab252690) and mouse anti tubulin (Abcam, Cat. ab7291), followed by goat anti-rabbit IgG IRDye® 800CW (Li-Cor, Cat. 926-32211) and goat anti-mouse IgG IRDye® 680RD (Li-Cor, Cat. 926-68070), respectively.	anti tubulin suggested: None
   Specific S protein expression was assessed via staining with Human anti SARS CoV S antibody (CR3022)	anti SARS CoV S suggested: None CR3022 suggested: None
   MRO-1214LC) followed by goat anti-human IgG F(ab’)2 fragment PE antibody (Immuno Research, Cat. 109-116-097) in a BD FACS Canto II cell analyzer and the FlowJo 10 software.	anti-human IgG suggested: (Jackson ImmunoResearch Labs Cat# 109-116-097, RRID:AB_2337677)
   Antibody analysis Blood samples were taken via retro-orbital bleeding and SARS-CoV-2 S -specific IgG1 and IgG2a antibodies (mice) or total IgG (hamster) antibodies were detected via ELISA.	IgG2a suggested: None total IgG suggested: None
   For detection, mouse sera were incubated with biotin rat anti-mouse IgG1 (BD Pharmingen, Cat. 550331) or biotin rat anti-mouse IgG2a (BD Pharmingen, Cat. 550332), hamster sera with biotin goat anti-hamster (Syrian) IgG antibody (BioLegend, Cat: 405601) followed by incubation with HRP-Streptavidin (BD, Cat: 554066).	anti-mouse IgG1 suggested: None anti-mouse IgG2a suggested: None anti-hamster ( Syrian ) IgG suggested: None
   The cells were then incubated with 200 ng/well of R-PE labelled rabbit anti-SARS-CoV-2 spike neutralizing monoclonal antibody (Sino Biological, Cat: 40592-R001) at 4°C for 2 h.	anti-SARS-CoV-2 suggested: (Abcam Cat# ab272854, RRID:AB_2847844)
   Antibody labelling with R-PE using the ZenonTM labelling kit for rabbit IgG (Invitrogen, Cat: Z25355) was performed according to the manufacturer’s instructions.	rabbit IgG suggested: None
   The following antibodies were used for flow cytometry analysis: anti-Thy1.2 FITC (clone 53-2.1; Biolegend, Cat.14304)	anti-Thy1.2 FITC suggested: (Abcam Cat# ab41584, RRID:AB_778437)
   Each dot represents an individual animal, bars depict the median. SECD: S ectodomain, RBD: receptor binding domain of S, NTD: N-terminal domain of S, nAb: neutralising antibody A ✱✱ C total IgG endpoint titer [Log10] d28 d42	Log10 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Quadruplicate, 10-fold serial dilutions were transferred to 96 well plates with Vero E6 cell culture monolayers and incubated for one hour at 37°C.	Vero E6 suggested: None
   HeLa cells were transfected with 2 µg of the mRNA component of CVnCoV.	HeLa suggested: CLS Cat# 300194/p772_HeLa, RRID:CVCL_0030)
   Experimental Models: Organisms/Strains
   Sentences	Resources
   For studies performed externally, Balb/c mice were provided and handled by Preclinics Gesellschaft für präklinische Forschung mbH, (Potsdam, Germany).	Balb/c suggested: RRID:IMSR_ORNL:BALB/cRl)
   Bu ffe r ad + ot ei n pr C Vn C oV µg B D d35 d49 IgG1 endpoint titer 9 d28 8 6 5 4 3 2 1 0 9 8 7 6 5 4 3 2 1 0 1st vacc d0 d7 d14d21 d0 d0 2nd vacc none + d0 d7 d14d21 d0 d0 2 d28	1st vacc d0 d7 d14d21 d0 d0 2nd vacc none + d0 d7 d14d21 d0 d0 2 suggested: None
   CVnCoV 2µg C d0 d7 d14d21 d0 d0 d28 protein + adj.	C d0 d7 d14d21 d0 d0 d28 protein + adj suggested: None
   IFN+/TNF+(% of CD4 T cells) IgG2a endpoint titer CD4 T-cells ✱✱ 0.8 0.6 0.4 0.2 0.0 d0 d7 d14 d21 d7 d7 + buffer CD8 T-cells 1st vacc 2nd vacc d28 d0 d7 d14 d21 d7 d7 none d35 d0 d7 d14 d21 d7 d7 d28 CVnCoV 2µg	d7 d14 d21 d7 d7 suggested: None vacc d28 d0 d7 d14 d21 d7 d7 none d35 d0 d7 d14 d21 d7 d7 d28 suggested: None
   d0 d7 d14 d21 d7 d7 d28 protein + adj. buffer IFN+ TNF+(% of CD8 T cells) VNT ✱✱✱ ✱✱ ✱✱ ✱ d0 d7 d14 d21 d7 d7 CVnCoV 2µg protein + adj. buffer Figure 2: CVnCoV elicits high levels of humoral and cellular immune responses.	d0 d7 d14 d21 d7 d7 d28 protein + adj suggested: None VNT ✱✱✱ ✱✱ ✱✱ ✱ d0 d7 d14 d21 d7 d7 CVnCoV 2µg protein + adj suggested: None
   ✱✱ d56 ✱✱ Throat swab titration [Log10 TCID50/ml] d5 d5 d5 d5 d6 d5 d5 d5 d5 d6 d5 d5 d5 d5 d6 untr.	Log10 TCID50/ml ] d5 d5 d5 d5 d6 d5 d5 d5 d5 d6 d5 d5 d5 d5 d6 suggested: None
   Software and Algorithms
   Sentences	Resources
   Flow cytometry data were analyzed using FlowJo software (Tree Star, Inc, Ashland, USA.).	FlowJo suggested: (FlowJo, RRID:SCR_008520)

   ---

   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.

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   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.

   Read the original source
4. 

   Version published to 10.1101/2020.10.23.351775 on bioRxiv

   Oct 23, 2020