Durability of mRNA-1273-induced antibodies against SARS-CoV-2 variants

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   ScreenIT

   May 21, 2021

   SciScore for 10.1101/2021.05.13.444010: (What is this?)

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

   Table 1: Rigor

   Ethics	Consent: Enrolled adults were healthy and provided informed consent prior to any study procedures.
   Sex as a biological variable	not detected.
   Randomization	8 subjects each were randomly chosen from participants from age cohorts 18-55, 56-70, and 71+ years of age who received two doses of 100 mcg mRNA-1273.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Cells were incubated with an anti-SARS-CoV spike primary antibody directly conjugated to biotin (CR3022-biotin) for 1 hour at room temperature.	anti-SARS-CoV spike suggested: None CR3022-biotin suggested: None
   Plates are washed and Sulfo-tag labeled anti IgG antibody is …

   SciScore for 10.1101/2021.05.13.444010: (What is this?)

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

   Table 1: Rigor

   Ethics	Consent: Enrolled adults were healthy and provided informed consent prior to any study procedures.
   Sex as a biological variable	not detected.
   Randomization	8 subjects each were randomly chosen from participants from age cohorts 18-55, 56-70, and 71+ years of age who received two doses of 100 mcg mRNA-1273.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Cells were incubated with an anti-SARS-CoV spike primary antibody directly conjugated to biotin (CR3022-biotin) for 1 hour at room temperature.	anti-SARS-CoV spike suggested: None CR3022-biotin suggested: None
   Plates are washed and Sulfo-tag labeled anti IgG antibody is applied to the wells and allowed to associate with complexed coated antigen – sample antibody within the assay wells.	anti IgG suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Cells and Viruses: VeroE6 cells were obtained from ATCC (clone E6, ATCC, #CRL-1586) and cultured in complete DMEM medium consisting of 1x DMEM (VWR, #45000-304), 10% FBS, 25mM HEPES Buffer (Corning Cellgro), 2mM L-glutamine, 1mM sodium pyruvate, 1x Non-essential Amino Acids, and 1x antibiotics.	VeroE6 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
   VeroE6-TMPRSS2 cells were kindly provided by Drs.	VeroE6-TMPRSS2 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
   Viruses were propagated in Vero-TMPRSS2 cells to generate viral stocks.	Vero-TMPRSS2 suggested: JCRB Cat# JCRB1818, RRID:CVCL_YQ48)
   Pseudoviruses were mixed with serial dilutions of sera or antibodies and then added to monolayers of ACE2-overexpressing 293T cells (gift of Michael Farzan and Huihui Mu), in triplicate.	293T suggested: CCLV Cat# CCLV-RIE 1018, RRID:CVCL_0063)
   Saponin [Sigma, 47036-250G-F] in PBS), was added to the fixed Vero cells for 20 minutes.	Vero suggested: None
   Cell-surface spike binding: HEK293T cells were transiently transfected with plasmids encoding full length SARS-CoV-2 spike variants using lipofectamine 3000 (L3000-001, ThermoFisher) following manufacturer’s protocol.	HEK293T suggested: None
   Recombinant DNA
   Sentences	Resources
   To produce SARS-CoV-2 pseudoviruses, an expression plasmid bearing codon-optimized SARS-CoV-2 full-length S plasmid was co-transfected into HEK293T/17 cells (ATCC#CRL-11268) cells with packaging plasmid pCMVDR8.2, luciferase reporter plasmid pHR′CMV-Luc (40) and a TMPRSS2 plasmid (41).	pCMVDR8.2 , luciferase reporter suggested: None pHR′CMV-Luc suggested: None TMPRSS2 suggested: RRID:Addgene_53887)
   Software and Algorithms
   Sentences	Resources
   All calculations are performed within Excel and the GraphPad Prism software, version 7.0.	Excel suggested: None GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)
   The samples were then acquired on a BD LSR Fortessa X-50 flow cytometer (BD biosciences) and analyzed using Flowjo (BD biosciences).	Flowjo suggested: (FlowJo, RRID:SCR_008520)

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

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

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2. 

   Version published to 10.1101/2021.05.13.444010v1 on bioRxiv

   May 16, 2021