RelCoVax®, a two antigen subunit protein vaccine candidate against SARS-CoV-2 induces strong immune responses in mice

Version published to 10.1016/j.vaccine.2022.06.026

Jul 1, 2022

SciScore for 10.1101/2022.01.07.475330: (What is this?)

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

Table 1: Rigor

Ethics	IACUC: Animal ethics, handling and treatment: Adult male and female BALB/c mice (inbred) were housed at Laboratory Animal Research Services, Reliance Life Sciences, Navi Mumbai.
Sex as a biological variable	Animal ethics, handling and treatment: Adult male and female BALB/c mice (inbred) were housed at Laboratory Animal Research Services, Reliance Life Sciences, Navi Mumbai.
Randomization	A total of 4 different studies were performed and animals in the studies were initially randomized into different groups based on body weight.
Blinding	not detected.
Power Analysis	not detected.
Cell Line Authentication	not detected.

Table 2: Resources

Antibodies
Sentences	Resources
General materials: In western blots, …

SciScore for 10.1101/2022.01.07.475330: (What is this?)

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

Table 1: Rigor

Ethics	IACUC: Animal ethics, handling and treatment: Adult male and female BALB/c mice (inbred) were housed at Laboratory Animal Research Services, Reliance Life Sciences, Navi Mumbai.
Sex as a biological variable	Animal ethics, handling and treatment: Adult male and female BALB/c mice (inbred) were housed at Laboratory Animal Research Services, Reliance Life Sciences, Navi Mumbai.
Randomization	A total of 4 different studies were performed and animals in the studies were initially randomized into different groups based on body weight.
Blinding	not detected.
Power Analysis	not detected.
Cell Line Authentication	not detected.

Table 2: Resources

Antibodies
Sentences	Resources
General materials: In western blots, RBD was detected using SARS-CoV-2 Spike RBD antibody (Prosci # 9087) as primary antibody and goat anti-rabbit alkaline phosphatase (Jackson Immunoresearch # 111-055-003) as secondary antibody.	anti-rabbit suggested: (Jackson ImmunoResearch Labs Cat# 111-055-003, RRID:AB_2337947)
N protein was detected using SARS-CoV-2 nucleocapsid antibody (Prosci # 35-579) as primary antibody and goat anti-mouse IgG whole molecule (Sigma # A4416) as secondary antibody.	anti-mouse IgG whole molecule suggested: (Sigma-Aldrich Cat# A4416, RRID:AB_258167)
Estimation of anti-RBD and anti-N antibodies in mouse sera by ELISA: Anti-RBD and anti-N antibodies were estimated in the sera of immunized mice by using the end-point titer ELISA method.	anti-RBD suggested: None anti-N suggested: None ELISA suggested: None
Experimental Models: Cell Lines
Sentences	Resources
DNA sequences for S and N proteins were codon optimized for expression in Cricetulus griseus (Chinese hamster) ovary or CHO cells and E. coli respectively and synthesized by GeneArt (Thermofisher Scientific).	CHO suggested: CLS Cat# 603479/p746_CHO, RRID:CVCL_0213)
After incubation, these were transferred in duplicate to the 24 well plate containing Vero cells after removal of growth media and incubated for 1 hr at 37°C and 5% CO2 with mixing every 15 mins for viral adsorption.	Vero suggested: None
Experimental Models: Organisms/Strains
Sentences	Resources
Animal ethics, handling and treatment: Adult male and female BALB/c mice (inbred) were housed at Laboratory Animal Research Services, Reliance Life Sciences, Navi Mumbai.	BALB/c suggested: RRID:IMSR_ORNL:BALB/cRl)
Recombinant DNA
Sentences	Resources
Similarly, the DNA sequence for the synthetic N gene (Met1 to Ala 419) was amplified and cloned into the NdeI and XhoI digested pET24a(+) vector (Novagen).	pET24a(+ suggested: None
The pXC17.4-RBD plasmid was transfected into CHOK1SV GS-KO cells (Lonza) by electroporation and stably transfected clones were selected by addition of methionine sulphoxamine (Sigma Aldrich) to the CD-CHO media (Gibco).	pXC17.4-RBD suggested: None
The pET24a(+)-N plasmid was transformed into chemically competent BL21DE3 pLysS E. coli cells (Novagen) and the transformants were isolated on kanamycin containing LB plates.	pET24a(+)-N suggested: None
Software and Algorithms
Sentences	Resources
After incubation at 37 °C for 1 hr, plates were washed 3 times with PBS-T and TMB substrate (Denovo Biolabs) was added for development of color.	Denovo Biolabs suggested: None
The plates were imaged using ImmunoSpot reader (Cellular Technologies Ltd) and number of spots per well were counted using Biospot 5.0 software. 2.10. Data Analysis: All data was analyzed and graphically represented by using Graphpad prism (9.2.0).	Graphpad suggested: (GraphPad, RRID:SCR_000306)
All preliminary data was tabulated and basic calculations were performed using Microsoft Excel.	Microsoft Excel suggested: (Microsoft Excel, RRID:SCR_016137)

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.
No funding statement was detected.
No protocol registration statement was detected.

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

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RelCoVax®, a two antigen subunit protein vaccine candidate against SARS-CoV-2 induces strong immune responses in mice

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