Distinct antibody responses to SARS-CoV-2 in children and adults across the COVID-19 clinical spectrum

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1. Version published to 10.1038/s41590-020-00826-9

   Nov 5, 2020
2. 

   ScreenIT

   Jul 14, 2020

   SciScore for 10.1101/2020.07.12.20151068: (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 Institutional Review Board at Columbia University Irving Medical Center. Consent: Written consent was obtained from CPD subjects.
   Randomization	not detected.
   Blinding	Cytopathic effect was revealed by crystal violet staining, and scored by an observer blinded to the study design and sample identity.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Peroxidase affiniPure goat anti-human IgG (H+L) antibody, anti-human IgM antibody (Jackson Immune Research, New Grove, PA)	anti-human IgG suggested: None anti-human IgM suggested: None
   , or …

   SciScore for 10.1101/2020.07.12.20151068: (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 Institutional Review Board at Columbia University Irving Medical Center. Consent: Written consent was obtained from CPD subjects.
   Randomization	not detected.
   Blinding	Cytopathic effect was revealed by crystal violet staining, and scored by an observer blinded to the study design and sample identity.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Peroxidase affiniPure goat anti-human IgG (H+L) antibody, anti-human IgM antibody (Jackson Immune Research, New Grove, PA)	anti-human IgG suggested: None anti-human IgM suggested: None
   , or anti-human IgA antibody (Thermofisher, Waltham, MA) was subsequently added into each well and incubated for 1hr at 37°C, washed and TMB substrate (Sigma, St. Louis) was added and the reaction was stopped using 1M sulfuric acid.	anti-human IgA suggested: None 1M sulfuric acid. suggested: None
   Quantitation of antibody titrations in ELISA and neutralization assays: For quantitation of neutralization titers in the pseudovirus assay, RFP signal driven by the pseudovirus normalized to the GFP signal derived from the SARS-Cov-2 S protein and ACE2 transfected cells was measured at 24 and 48hr; the ratio of normalized RFP at 48 hours (RFP48) to normalized RFP at 24 hours (RFP24) was calculated.	ACE2 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   This expression vector was transiently transfected into HEK293F cells and the spike trimer secreted in the supernatant was purified 3-5 days post transfection by metal affinity chromatography using an Ni-NTA (Qiagen) column.	HEK293F suggested: None
   Initially, VSV-ΔG-RFP pseudotyped with VSV G is used to infect 293T (human kidney epithelial) cells that were co-transfected with full length codon optimized SARS-CoV-2 S-protein (Epoch Life Science, Sugar Land, TX), the viral entry receptor ACE2 (Epoch Life Science, Sugar Land, TX) and green fluorescent protein (GFP).	293T suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
   During the assay, infected, S protein-expressing HEK293T cells generate VSV-ΔG-RFP viruses that bear S protein which subsequently infects and drives RFP expression in Vero cells and undergo multiple cycles of entry and budding in the HEK293T cells due to the co-expression of S protein with ACE2.	Vero suggested: None HEK293T suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
   Live virus neutralization assay: Two-fold dilutions of plasma in 50µL of Dulbecco’s Modified Eagle Media (DMEM) were incubated with 200 plaque forming unit (pfu) of SARS-CoV-2 in 50µL of DMEM for 30min at 4°C. 100µL of DMEM 4%FBS containing 4×104 Vero E6 cells were added on the top of the former mix in order to have final dilution of sera from 1:50 to 1:6400 (4 wells per dilution).	Vero E6 suggested: None
   Software and Algorithms
   Sentences	Resources
   Image analysis was performed using ImageJ 36.	ImageJ suggested: (ImageJ, RRID:SCR_003070)
   Heatmaps of this ratio at all titrations for all samples were generated using the Python data visualization library, seaborn41	Python suggested: (IPython, RRID:SCR_001658)
   Statistical analysis: All statistical analysis was performed using Prism 8 software (GraphPad, San Diego, CA).	Prism suggested: (PRISM, RRID:SCR_005375) GraphPad 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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3. 

   Version published to 10.1101/2020.07.12.20151068v1 on medRxiv

   Jul 14, 2020