Elicitation of potent SARS-CoV-2 neutralizing antibody responses through immunization with a versatile adenovirus-inspired multimerization platform

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1. Version published to 10.1016/j.ymthe.2022.02.011

   May 1, 2022
2. 

   ScreenIT

   Sep 15, 2021

   SciScore for 10.1101/2021.09.13.460076: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: In vivo experiments and vaccination: Vaccination experiments have been performed according to ethical guidelines, under a protocol approved by the Grenoble Ethical Committee for Animal Experimentation and the French Ministry of Higher Education and Researcher (reference number: APAFIS#27765-2020102114206782 v2).
   Sex as a biological variable	5-week-old female Balb/c mice were purchased from Janvier (Le Genestet St. Isle, France).
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Experimental Models: Cell Lines
   Sentences	Resources
   EXPI293 cells grown in EXPI293 expression medium were transiently transfected with …

   SciScore for 10.1101/2021.09.13.460076: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: In vivo experiments and vaccination: Vaccination experiments have been performed according to ethical guidelines, under a protocol approved by the Grenoble Ethical Committee for Animal Experimentation and the French Ministry of Higher Education and Researcher (reference number: APAFIS#27765-2020102114206782 v2).
   Sex as a biological variable	5-week-old female Balb/c mice were purchased from Janvier (Le Genestet St. Isle, France).
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Experimental Models: Cell Lines
   Sentences	Resources
   EXPI293 cells grown in EXPI293 expression medium were transiently transfected with the vector according to the manufacturer’s protocol (Thermo Fisher Scientific).	EXPI293 suggested: RRID:CVCL_D615)
   Briefly, gag/pol and luciferase plasmids were co-transfected with a SARS-CoV-2 spike plasmid with a C-term deletion of 18aa at 1/0,4/1 ratio on adherent HEK293T cells.	HEK293T suggested: CCLV Cat# CCLV-RIE 1018, RRID:CVCL_0063)
   Before use, supernatants were tittered using HeLa ACE-2 cells to determine the appropriate dilution of pseudovirus necessary to obtain about 150 000 RLU per well in a 48 well-plate.	HeLa ACE-2 suggested: None
   Neutralization assay (mAb): Serial 3-fold dilutions starting from 1/10 dilution (serum), or 6µg/mL (known bNAbs) were let in contact with the pseudoviruses for 1h at 37°C in 96-w white plates (Greiner #675083), before addition of HeLa ACE2 cells.	HeLa ACE2 suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   5-week-old female Balb/c mice were purchased from Janvier (Le Genestet St. Isle, France).	Balb/c suggested: RRID:IMSR_ORNL:BALB/cRl)
   Recombinant DNA
   Sentences	Resources
   Synthetic DNA (Genscript) was cloned in the pACEBac1 using the restriction sites BamHI and HindIII.	pACEBac1 suggested: RRID:Addgene_109156)
   RBD purification: The following reagent was produced under HHSN272201400008C and obtained through BEI Resources, NIAID, NIH: Vector pCAGGS Containing the SARS-Related Coronavirus 2, Wuhan-Hu-1 Spike Glycoprotein Receptor Binding Domain (RBD), NR-52309.	pCAGGS suggested: RRID:Addgene_18926)
   Software and Algorithms
   Sentences	Resources
   Automated data collection was performed with SerialEM, acquiring one image per hole, in counting mode.	SerialEM suggested: (SerialEM, RRID:SCR_017293)
   Image processing and cryo-EM structure refinement: Movie drift correction was performed with Motioncor2 37 using frames from 2 to 40.	Motioncor2 suggested: (MotionCor2, RRID:SCR_016499)
   CTF determination was performed with Relion 3.1.2 (Scheres 2012).	Relion suggested: (RELION, RRID:SCR_016274)
   Statistical analyses: For ELISA and neutralization assays, IC50 were deduced from crude data after normalization using GraphPad Prism “log(inhibitor) vs normalized response” function.	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 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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3. 

   Version published to 10.1101/2021.09.13.460076 on bioRxiv

   Sep 13, 2021