RelCoVax®, a two antigen subunit protein vaccine candidate against SARS-CoV-2 induces strong immune responses in mice
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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-rabbitsuggested: (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 moleculesuggested: (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-RBDsuggested: Noneanti-Nsuggested: NoneELISAsuggested: NoneExperimental 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). CHOsuggested: 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. Verosuggested: NoneExperimental 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/csuggested: 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: NoneThe 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-RBDsuggested: NoneThe 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(+)-Nsuggested: NoneSoftware 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 Biolabssuggested: NoneThe 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). Graphpadsuggested: (GraphPad, RRID:SCR_000306)All preliminary data was tabulated and basic calculations were performed using Microsoft Excel. Microsoft Excelsuggested: (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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