Neutralizing antibody against SARS-CoV-2 spike in COVID-19 patients, health care workers, and convalescent plasma donors

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1. 

   ScreenIT

   Mar 1, 2021

   SciScore for 10.1101/2020.08.02.20166819: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   Randomization	not detected.
   Blinding	Plasma and serum were collected from hospitalized COVID-19 inpatients or ICU patients, OSU health care workers, and blinded convalescent plasma donors and analyzed in a blinded manner.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Antibodies used for Western blotting included anti-C9 (anti-rhodopsin) (Santa Cruz, #57432), anti-p24 (abcam, #ab63917), anti-β-actin (Sigma, #A1978) and secondary antibodies anti-Mouse IgG (Sigma, #A5278)	anti-C9 suggested: None anti-C9 ( anti-rhodopsin ) suggested: None anti-p24 suggested: …

   SciScore for 10.1101/2020.08.02.20166819: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   Randomization	not detected.
   Blinding	Plasma and serum were collected from hospitalized COVID-19 inpatients or ICU patients, OSU health care workers, and blinded convalescent plasma donors and analyzed in a blinded manner.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Antibodies used for Western blotting included anti-C9 (anti-rhodopsin) (Santa Cruz, #57432), anti-p24 (abcam, #ab63917), anti-β-actin (Sigma, #A1978) and secondary antibodies anti-Mouse IgG (Sigma, #A5278)	anti-C9 suggested: None anti-C9 ( anti-rhodopsin ) suggested: None anti-p24 suggested: None anti-β-actin suggested: None anti-Mouse IgG suggested: (Sigma-Aldrich Cat# A5278, RRID:AB_258232)
   Secondary antibodies used for flow cytometry included FITC-conjugated anti-human IgG-Fc (Sigma,405 #F9512).	anti-human IgG-Fc suggested: None
   After three washes with PBS plus 2% FBS, cells were incubated with FITC-conjugated anti-human IgG (1:200, Sigma, #F0257) secondary antibodies for 1 hr.	anti-human IgG suggested: None
   Following wash steps, wells were treated with 100 μl of HRP labeled anti-human-IgG tracer antibody (EDI, #31220), incubated at room temperature for 30 min, and again washed.	anti-human-IgG tracer suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   HEK293T (ATCC CRL-11268, RRID: CVCL_1926), HeLa (ATCC, CCL-2, RRID: CVCL_0030), HTX (a subclone of HT1080), A549 (ATCC, CCL-185, RRID: CVCL_0023) and Huh7.5 (RRID: CVCL_7927) cells were grown in Dulbecco’s modified Eagle’s medium (DMEM), supplemented with 1% penicillin/streptomycin and 10% (vol/vol) FBS.	HEK293T detected: (ATCC Cat# CRL-11268, RRID:CVCL_1926) HeLa detected: (BCRC Cat# 60005, RRID:CVCL_0030) HT1080 suggested: None A549 suggested: None HTX a detected: (BCRC Cat# 60074, RRID:CVCL_0023) Huh7.5 detected: ( RRID:CVCL_7927)
   Calu-3 (ATCC, gift of Stelle Cormet-Boyaka) were grown in Eagle’s Minimum Essential Medium (EMEM), supplemented with 1% penicillin/streptomycin and 10% (vol/vol) FBS.	Calu-3 suggested: KCLB Cat# 30055, RRID:CVCL_0609)
   HEK293T/ACE2 cell line is a gift from Dr. Fang Li at the University of Minnesota. HeLa, A549, HTX and Huh7.5 cells stably expressing ACE2 were generated by transduction of pLenti-GFP vectors expressing ACE2 (OriGene, #RC208442L4), followed by puromycin selection (1 μg/mL) for 6 days.	HEK293T/ACE2 suggested: None HeLa suggested: None Huh7.5 suggested: RRID:CVCL_7927)
   Production of intron-Gluc- or secNluc-based lentiviral pseudotypes bearing the S protein of SARS Coronaviruses and viral infection: For intron-Gluc- or secNluc-based pseudotyped lentiviral production, we transfected HEK293T cells with HIV-1-NL4.3 inGluc or secNluc vector plus a plasmid expressing the S protein or VSV-G in a 2:1 ratio using polyethylenimine (PEI).	HEK293T suggested: None
   Serum/virus mixtures were then used to infect confluent Vero-E6 cells for 1 h at 37°C.	Vero-E6 suggested: None
   Software and Algorithms
   Sentences	Resources
   Statistical analyses were performed using GraphPad Prism 5.0 as follows: One-way Analysis of Variance (ANOVA) with Bonferroni’s post-tests was used to compute statistical significance between multiple groups for multiple comparison.	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 found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

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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 10.1172/jci.insight.143213

   Nov 19, 2020
3. 

   Version published to 10.1101/2020.08.02.20166819 on medRxiv

   Aug 4, 2020