Engineering, production and characterization of Spike and Nucleocapsid structural proteins of SARS–CoV-2 in Nicotiana benthamiana as vaccine candidates against COVID-19

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   ScreenIT

   Dec 31, 2020

   SciScore for 10.1101/2020.12.29.424779: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: Mice studies were carried out at Akdeniz University Experimental Animal CareUnit under permission of the Local Ethics Committee for Animal Experiments at Akdeniz University with thesupervision of a veterinarian.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   After transfer, Western blot membranes were blocked with I-Block (Applied Biosystems, Carlsbad, CA) and recombinant proteins N, RBD, S1 and N+RBD were detected with anti-DYKDDDDK antibody (cat. no. 651503, BioLegend, for N, RBD or N+RBD),	anti-DYKDDDDK suggested: None
   anti-His antibody (for …

   SciScore for 10.1101/2020.12.29.424779: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: Mice studies were carried out at Akdeniz University Experimental Animal CareUnit under permission of the Local Ethics Committee for Animal Experiments at Akdeniz University with thesupervision of a veterinarian.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   After transfer, Western blot membranes were blocked with I-Block (Applied Biosystems, Carlsbad, CA) and recombinant proteins N, RBD, S1 and N+RBD were detected with anti-DYKDDDDK antibody (cat. no. 651503, BioLegend, for N, RBD or N+RBD),	anti-DYKDDDDK suggested: None
   anti-His antibody (for S1) and anti-RBD of S protein of SARC-CoV-2 monoclonal antibody (cat. no. MBS7135930, for RBD) or Human Novel Coronavirus Nucleoprotein (N) (1-419aa) monoclonal Antibody (MyBioSource, cat. no. MBS7135930).	1-419aa suggested: None
   After washing, wells were incubated with anti-rabbit IgG antibody (Cat. no. MBS440123, MyBioSource, USA).	anti-rabbit IgG suggested: (MyBioSource Cat# MBS440123, RRID:AB_10571491)
   Serum samples were collected on days −1 (pre-bleed) and 42 (post 2nd vaccination) and assessed for anti-N, anti-RBD, anti-N+RBD and anti-S1 domain antibody responses by an IgG ELISA.	anti-N suggested: None anti-RBD suggested: None anti-N+RBD suggested: None anti-S1 domain suggested: None
   After transfer, Western blot membranes were blocked with I-Block (Applied Biosystems, Carlsbad, CA) and recombinant proteins detected with an anti-FLAG (N, S1 and N+S1) or anti-His Antibody (S2), or anti-SARS-COV2 COVID 19 Spike Protein Coronavirus Monoclonal Antibody (MyBioSource, cat. no. MBS2563837).	anti-FLAG suggested: None anti-His suggested: None anti-SARS-COV2 COVID 19 Spike Protein Coronavirus suggested: None
   Software and Algorithms
   Sentences	Resources
   Total protein content was estimated using the BioDrop and then analyzed by SDS-PAGE and western blot.	BioDrop suggested: None
   The degradation of gRBD, dRBD or gS1 protein bands were calculated and quantified based on SDS-PAGE and WB analysis using highly sensitive Gene Tools software (Syngene Bioimaging, UK) and ImageJ software as described previously21.	ImageJ suggested: (ImageJ, RRID:SCR_003070)
   GraphPad Prism software was used for all statistical analyses.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)

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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.

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   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).

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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.

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2. 

   Version published to 10.1101/2020.12.29.424779v1 on bioRxiv

   Dec 30, 2020