Preclinical evaluation of a COVID-19 vaccine candidate based on a recombinant RBD fusion heterodimer of SARS-CoV-2
This article has been Reviewed by the following groups
Discuss this preprint
Start a discussion What are Sciety discussions?Listed in
- Evaluated articles (ScreenIT)
Abstract
No abstract available
Article activity feed
-
-
SciScore for 10.1101/2021.11.22.469117: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: All procedures that involved BALB/c mice were conducted in accordance with the European Union Guidelines for Animal Welfare (Directive 2010/63/EU) and approved by the Ethics Committee of HIPRA Scientific S.L.U. and the Department of Territori i Sostenibilitat of the Catalan Government (file: 11388). Sex as a biological variable Viral RNA was determined by quantitative real-time quantitative polymerase chain reaction (RT-qPCR) at D37 (2 days postinfection), D39 (4 days post-infection), D42 (in males, 7 days post-infection) and D43 (in females, 8 days post-infection), or at the time of euthanasia in animals reaching end-point criteria before the scheduled euthanasia day. Randomization not … SciScore for 10.1101/2021.11.22.469117: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: All procedures that involved BALB/c mice were conducted in accordance with the European Union Guidelines for Animal Welfare (Directive 2010/63/EU) and approved by the Ethics Committee of HIPRA Scientific S.L.U. and the Department of Territori i Sostenibilitat of the Catalan Government (file: 11388). Sex as a biological variable Viral RNA was determined by quantitative real-time quantitative polymerase chain reaction (RT-qPCR) at D37 (2 days postinfection), D39 (4 days post-infection), D42 (in males, 7 days post-infection) and D43 (in females, 8 days post-infection), or at the time of euthanasia in animals reaching end-point criteria before the scheduled euthanasia day. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources RBD-specific binding antibody titres The mouse strain B6.Cg-Tg(K18-ACE2)2Prlmn/J (K18-hACE2) (Jackson Laboratories, ME, USA) and BALB/c (Envigo, IN, USA) were immunized with different doses of the recombinant RBD fusion heterodimer antigen. 2.2.2. 2.2.3. (K18-hACE2)suggested: NoneK18-hACE2suggested: NoneWells were incubated with serial dilutions of the serum samples and the bound total IgG specific antibodies were detected by Peroxidase-conjugated Goat Anti-Mouse IgG (Sigma-Aldrich). Anti-Mouse IgGsuggested: NoneFor flow cytometric analysis, equal numbers of cells were stained with fixable viability stain 780 (BD Biosciences, New Jersey, NJ, USA) in PBS for 15 min at RT followed by staining with antibodies against CD3, CD4, CD8 and CD44 for 20 min on ice in FACS buffer (PBS 5% FCS, 0.5% BSA, 0.07% NaN3). CD3suggested: (Abcam Cat# ab52305, RRID:AB_955118)CD4suggested: (RayBiotech Cat# CS-11-0133, RRID:AB_1228052)CD8suggested: (RayBiotech Cat# CS-11-0250, RRID:AB_1228199)CD44suggested: NoneCells were then fixed for 20 min on ice with 2% Formaldehyde and stained with antibodies against intracellular proteins (IFNγ, TNFα, IL-2 and IL-4) for 20 min on ice in perm/wash buffer (PBS 1% FCS, NaN3 0.1%, Saponin 0.1%). TNFαsuggested: NoneIL-4suggested: NoneExperimental Models: Cell Lines Sentences Resources For the neutralization assay, 200 TCID50 of pseudovirus supernatant was preincubated with serial dilutions of the heat-inactivated serum samples for 1 h at 37 °C and then added onto ACE2 overexpressing HEK293T cells. HEK293Tsuggested: KCB Cat# KCB 200744YJ, RRID:CVCL_0063)Virus titration Virus titres were determined using a standard TCID50 assay in Vero E6 cells at CReSA (IRTA-UAB), according to the method described by Brustolin et al., 2021. Vero E6suggested: NoneExperimental Models: Organisms/Strains Sentences Resources RBD-specific binding antibody titres The mouse strain B6.Cg-Tg(K18-ACE2)2Prlmn/J (K18-hACE2) (Jackson Laboratories, ME, USA) and BALB/c (Envigo, IN, USA) were immunized with different doses of the recombinant RBD fusion heterodimer antigen. 2.2.2. 2.2.3. B6.Cg-Tg(K18-ACE2)2Prlmn/J (K18-hACE2)suggested: NoneAnimal study design BALB/c mice (Envigo) and transgenic B6.Cg-Tg(K18-ACE2)2Prlmn/J (K-18-hACE2) (Jackson Laboratories) were used as animal models. B6.Cg-Tg(K18-ACE2)2Prlmn/J (K-18-hACE2)suggested: NoneAll procedures that involved BALB/c mice were conducted in accordance with the European Union Guidelines for Animal Welfare (Directive 2010/63/EU) and approved by the Ethics Committee of HIPRA Scientific S.L.U. and the Department of Territori i Sostenibilitat of the Catalan Government (file: 11388). BALB/csuggested: NoneSoftware and Algorithms Sentences Resources The affinity test of the RBD dimer with human ACE2 by surface plasmon resonance (SPR) was performed by ACROBiosystems. ACROBiosystemssuggested: (ACRObiosystems, RRID:SCR_012550)IC50 were calculated by plotting and fitting neutralization values and the lo fog plasma dilution to a 4-parameters equation in Prism 9.0.2 (GraphPad Software, USA). 4.7. Prismsuggested: (PRISM, RRID:SCR_005375)GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)FACS data were analysed using flowjo 10 software (Tree Star Inc., Ashland, OR, USA). flowjosuggested: (FlowJo, RRID:SCR_008520)Statistical analysis All statistical analyses were performed using R (version 4.0.5), SPSS (version 22), and/or GraphPad Prism (version 9). SPSSsuggested: (SPSS, RRID:SCR_002865)GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)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: We found the following clinical trial numbers in your paper:
Identifier Status Title NCT05007509 Active, not recruiting Safety and Immunogenicity Study of Recombinant Protein RBD C… 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.
- 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.
Results from scite Reference Check: We found no unreliable references.
-
