Neutralizing antibody and soluble ACE2 inhibition of a replication-competent VSV-SARS-CoV-2 and a clinical isolate of SARS-CoV-2

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   ScreenIT

   Jun 8, 2020

   SciScore for 10.1101/2020.05.18.102038: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: This study was approved by the Mayo Clinic Institutional Review
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Viral proteins were separated on a 8% acrylamide gel, transferred onto a nitrocellulose membrane, and incubated with human anti-SARS antibody CR3022 diluted in Tris-buffer saline containing 1% Tween-20 (TBS-T) and 5% milk, followed by incubation with HRP-conjugated goat anti-human antibody (Abcam) diluted in TBS-T containing 1% milk.	anti-SARS suggested: None anti-human antibody suggested: None
   Samples …

   SciScore for 10.1101/2020.05.18.102038: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: This study was approved by the Mayo Clinic Institutional Review
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Viral proteins were separated on a 8% acrylamide gel, transferred onto a nitrocellulose membrane, and incubated with human anti-SARS antibody CR3022 diluted in Tris-buffer saline containing 1% Tween-20 (TBS-T) and 5% milk, followed by incubation with HRP-conjugated goat anti-human antibody (Abcam) diluted in TBS-T containing 1% milk.	anti-SARS suggested: None anti-human antibody suggested: None
   Samples were prescreened by the Euroimmun anti-SARS-CoV-2 IgG ELISA (Lubeck, Germany), a qualitative assay with the Food and Drug Administration Emergency Use Authorization that detects antibodies to the SARS-CoV-2 S protein.	anti-SARS-CoV-2 IgG ELISA ( Lubeck , Germany) suggested: None
   Plates were washed and sequentially incubated with 1 μg/mL of CR3022 anti-S antibody (ter Meulen et al., 2006; Yuan et al., 2020) and HRP-conjugated goat anti-human IgG in PBS supplemented with 0.1% saponin and 0.1% BSA.	anti-S suggested: None anti-human IgG suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   BSRT7/5, Vero CCL81, Vero E6, Vero E6-TMPRSS2, A549, Caco-2, Caco-2 BBe1	A549 suggested: NCI-DTP Cat# A549, RRID:CVCL_0023) Caco-2 suggested: None
   , Huh7, HepG2, Hela, BHK-21, HEK293, and HEK293T were maintained in DMEM (Corning or VWR) supplemented with glucose, L-glutamine, sodium pyruvate, and 10% fetal bovine serum (FBS).	Huh7 suggested: None HepG2 suggested: None BHK-21 suggested: None HEK293 suggested: None
   HT-29 cells were cultured in complete DMEM/F12 (Thermo-Fisher) supplemented with sodium pyruvate, non-essential amino acids, and HEPES.	HT-29 suggested: None
   Vero E6-TMPRSS2 cells were generated using a lentivirus vector.	Vero E6-TMPRSS2 suggested: None
   Briefly, HEK293T producer cells were transfected with pLX304-TMPRSS2, pCAGGS-VSV-G, and psPAX2, and cell culture supernatants were collected at 48 hours and clarified by centrifugation at 1,000 x g for 5 min.	HEK293T suggested: None
   The resulting lentivirus was used to infect Vero E6 cells for 24 h, and cells were selected with 40 μg/ml blasticidin for 7 days.	Vero E6 suggested: None
   Briefly, BSRT7/5 cells were inoculated with vaccinia virus vTF7-3 and subsequently transfected with T7-expression plasmids encoding VSV N, P, L, and G, and an antigenomic copy of the viral genome.	BSRT7/5 suggested: CCLV Cat# CCLV-RIE 0583, RRID:CVCL_RW96)
   Next generation sequencing: Total RNA was extracted from Vero CCL81 cells infected with VSV-SARS-CoV-2-SΔ21 using Trizol (Invitrogen) according to the manufacturer’s protocol.	Vero CCL81 suggested: None
   Software and Algorithms
   Sentences	Resources
   Other plasmids were previously described: VSV N, P, L and G expression plasmids (Stanifer et al., 2011; Whelan et al., 1995), psPAX2 (Addgene), and pLX304-TMPRSS2 (Zang et al., 2020).	Addgene suggested: (Addgene, RRID:SCR_002037)
   Alignment of each sample to VSV and SARS CoV-2 sequence was performed using bbmap v38.79	bbmap suggested: (BBmap, RRID:SCR_016965)
   Mapped reads were extracted using samtools 1.9 and used for de novo assembly by SPAdes v3.13.0.	samtools suggested: (SAMTOOLS, RRID:SCR_002105) SPAdes suggested: (SPAdes, RRID:SCR_000131)
   Consensus sequences for each RNA sample were generated by aligning contigs to the reference plasmid sequence pVSV(1+)-eGFP-SARS-CoV-2-S with SnapGene v5.0.	SnapGene suggested: (SnapGene, RRID:SCR_015052)
   Data were processed using Prism software (GraphPad Prism 8.0).	Prism suggested: (PRISM, RRID:SCR_005375) GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)
   Data were processed using Prism software (GraphPad Prism 8.0).	Prism suggested: (PRISM, RRID:SCR_005375) GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)
   Statistical analyses: All statistical tests were performed using GraphPad Prism 8.0 software as described in the indicated figure legends.	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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2. 

   Version published to 10.1101/2020.05.18.102038 on bioRxiv

   May 18, 2020