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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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-SARSsuggested: Noneanti-human antibodysuggested: NoneSamples … 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-SARSsuggested: Noneanti-human antibodysuggested: NoneSamples 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: NonePlates 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-Ssuggested: Noneanti-human IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources BSRT7/5, Vero CCL81, Vero E6, Vero E6-TMPRSS2, A549, Caco-2, Caco-2 BBe1 A549suggested: NCI-DTP Cat# A549, RRID:CVCL_0023)Caco-2suggested: 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). Huh7suggested: NoneHepG2suggested: NoneBHK-21suggested: NoneHEK293suggested: NoneHT-29 cells were cultured in complete DMEM/F12 (Thermo-Fisher) supplemented with sodium pyruvate, non-essential amino acids, and HEPES. HT-29suggested: NoneVero E6-TMPRSS2 cells were generated using a lentivirus vector. Vero E6-TMPRSS2suggested: NoneBriefly, 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. HEK293Tsuggested: NoneThe 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 E6suggested: NoneBriefly, 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. BSRT7/5suggested: 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 CCL81suggested: NoneSoftware 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). Addgenesuggested: (Addgene, RRID:SCR_002037)Alignment of each sample to VSV and SARS CoV-2 sequence was performed using bbmap v38.79 bbmapsuggested: (BBmap, RRID:SCR_016965)Mapped reads were extracted using samtools 1.9 and used for de novo assembly by SPAdes v3.13.0. samtoolssuggested: (SAMTOOLS, RRID:SCR_002105)SPAdessuggested: (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. SnapGenesuggested: (SnapGene, RRID:SCR_015052)Data were processed using Prism software (GraphPad Prism 8.0). Prismsuggested: (PRISM, RRID:SCR_005375)GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)Data were processed using Prism software (GraphPad Prism 8.0). Prismsuggested: (PRISM, RRID:SCR_005375)GraphPadsuggested: (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 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: 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.
- 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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