PRE-CLINICAL IMMUNE RESPONSE AND SAFETY EVALUATION OF THE PROTEIN SUBUNIT VACCINE NANOCOVAX FOR COVID-19

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   ScreenIT

   Jul 26, 2021

   SciScore for 10.1101/2021.07.20.453162: (What is this?)

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

   Table 1: Rigor

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   Ethics

   Euthanasia Agents: At the end of the study, all mice were sacrificed by cervical dislocation and some vital organs, like kidneys, liver and spleen were collected to examine morphological and histological characteristics and weights of the organs. 2.6.2. IACUC: Animal ethics statement: This study was carried out in strict adherence to the animal laboratory of Nanogen pharmaceutical Company, the National Institute of drug quality control (NIDQC), and the laboratory of Hanoi Medical University (HMU). IRB: The processes were designed according to the guide of ICH/GCP, Drug administration of Vietnam, as well ACTD and approved by Ethics committee of Ministry of Health 2.8.

   Sex as a biological variable

   SciScore for 10.1101/2021.07.20.453162: (What is this?)

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

   Table 1: Rigor

   Ethics	Euthanasia Agents: At the end of the study, all mice were sacrificed by cervical dislocation and some vital organs, like kidneys, liver and spleen were collected to examine morphological and histological characteristics and weights of the organs. 2.6.2. IACUC: Animal ethics statement: This study was carried out in strict adherence to the animal laboratory of Nanogen pharmaceutical Company, the National Institute of drug quality control (NIDQC), and the laboratory of Hanoi Medical University (HMU). IRB: The processes were designed according to the guide of ICH/GCP, Drug administration of Vietnam, as well ACTD and approved by Ethics committee of Ministry of Health 2.8.
   Sex as a biological variable	Animal vaccination: Balb/c mice of both sexes, 6-10 weeks old were used for immunological studies.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Next, the membranes were blocked in 0.5% BSA (Sigma-Aldrich, St. Louis, MO, USA) and incubated with human anti-S1 antibody (Abcam, Cambridge, MA, UK) for 3 hours at room temperature.	anti-S1 suggested: None
   After washing, the membranes were incubated with horseradish peroxidase (HRP)-linked rabbit anti human IgG antibody (Abcam, Cambridge, MA, UK) for 1 hour at room temperature.	anti human IgG suggested: None
   The Balb/c mice, Syrian hamsters, as well non-human primates were immunized intramuscularly with Nanocovax at doses of 25 μg, 50 μg, 75 μg, and 100 μg, their serum samples were collected quantification of spike protein specific IgG antibodies by ELISA.	specific IgG suggested: None
   Upper serum fraction was collected and heat-inactivated at 56°C in 30 minutes before use or kept at −20°C. 2.4.2. Quantification of SARS-CoV-2 spike protein specific IgG antibody by ELISA: The method of quantifying SARS-CoV-2 spike protein-specific IgG antibody was adapted from Tan C. W. et al, [15].	SARS-CoV-2 spike protein specific IgG suggested: None SARS-CoV-2 spike protein-specific IgG suggested: None
   Wells were washed 3 times with washing buffer then 100 μL of diluted goat anti-mouse IgG (Fc specific)-Peroxidase antibody (#A0168; Sigma-aldrich) were added.	anti-mouse IgG ( Fc specific)-Peroxidase suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   The gene encoding the Spike protein of SARS-CoV-2 (UniProt P0DTC2) codon-optimized for expression in CHO cells was synthesized by Genscript (Piscataway, NJ, USA).	CHO suggested: CLS Cat# 603479/p746_CHO, RRID:CVCL_0213)
   The isolate was propagated in Vero E6 to prepare virus stocks.	Vero E6 suggested: RRID:CVCL_XD71)
   Experimental Models: Organisms/Strains
   Sentences	Resources
   In vitro surrogate virus neutralization assay: The virus neutralization ability of antibodies in sera of Balb/C mice, hamster, and non-human private were determined by the virus surrogate neutralization kit (cat # L00847, Genscript, Singapore).	Balb/C suggested: None
   Recombinant DNA
   Sentences	Resources
   The pCNeoMEM plasmid containes a G418 resistance gene used as a selection marker, a MoMLV promoter to express the target gene, a human universal chromatin opening element (UCOE), untranslated regions from the Chinese hamster EEF1A1 gene (eEF1A1), and a synthetic matrix attachment sequence (sMAR).	pCNeoMEM suggested: None
   The optimized DNA fragment was cloned in the expression vector to create pCNeoMEM-S, which was completely sequenced by NextGen technology at the Center for Computational and Integrative Biology, Harvard University. 2.1.2.	pCNeoMEM-S suggested: None
   Software and Algorithms
   Sentences	Resources
   Statistical analysis: The collected data were statistically analyzed using Grapthpad Prism, version 5 (Grapthpad Software).	Grapthpad Prism suggested: None

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   Results from OddPub: Thank you for sharing your data.

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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 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.

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   Results from scite Reference Check: We found no unreliable references.

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   Version published to 10.1101/2021.07.20.453162v1 on bioRxiv

   Jul 21, 2021