Comparative analysis of antigen-specific anti-SARS-CoV-2 antibody isotypes in COVID-19 patients

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1. 

   ScreenIT

   Dec 5, 2020

   SciScore for 10.1101/2020.12.04.407510: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: Samples: This study was reviewed and approved by the Ethics Committee for Clinical Research of the Center for Research Promotion and Support in Fujita Health University
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	One hundred serum samples obtained from 100 healthy human volunteers (mean age, 47; males, 58 and females, 42), collected before the COVID-19 pandemic (July 2012), were used as negative controls to evaluate the specificity and cut-off values for each assay.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The plates were washed three times with 300 μL PBST and …

   SciScore for 10.1101/2020.12.04.407510: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: Samples: This study was reviewed and approved by the Ethics Committee for Clinical Research of the Center for Research Promotion and Support in Fujita Health University
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	One hundred serum samples obtained from 100 healthy human volunteers (mean age, 47; males, 58 and females, 42), collected before the COVID-19 pandemic (July 2012), were used as negative controls to evaluate the specificity and cut-off values for each assay.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The plates were washed three times with 300 μL PBST and 50 μL of peroxidase-labelled anti-human IgG, IgM or IgA antibody was added per well and incubated at RT for 60 min.	anti-human IgG suggested: None IgA suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   ELISA development and assay: SARS-CoV-2 RBD, S1, S full, S trimer and N protein expressed in HEK293 cells were selected as the target antigens.	HEK293 suggested: CLS Cat# 300192/p777_HEK293, RRID:CVCL_0045)
   The mixture of diluted sera and 100 TCID50 SARS-CoV-2 JPN/TY/WK-521 strain were incubated at 37°C for 1 hour, then placed on VeroE6/TMRRSS2 cells (JCRB1819, JCRB Cell Bank) and cultured at 37°C with 5% CO2 (19).	VeroE6/TMRRSS2 suggested: None
   Software and Algorithms
   Sentences	Resources
   Statistical analysis: Statistical analysis was performed using GraphPad Prism version 8.0.0 for Windows (GraphPad Software, USA)	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798) 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: We detected the following sentences addressing limitations in the study:

   There are several limitations in this study. Importantly, there were only four severe COVID-19 patients from whom residual serum samples were available to investigate the association between serum RBD-IgG levels and neutralizing activity. Since these four severe COVID-19 patients all recovered from COVID-19, we could not investigate differences in antibody responses and neutralizing activity between patients who recovered from COVID-19 and those who did not. In summary, our results indicate that the anti-SARS-CoV-2 antibody response in COVID-19 patients varies among the antigen-specific antibody isotypes. Diagnostic performance of ELISAs for detection of anti-SARS-CoV-2 antibodies also varies among antigen-specific antibody isotypes. Among them, serum RBD-IgG levels best correlate with virus neutralizing activity and disease severity, thus may be the optimal assay to track COVID-19 seroconversion responses and use as the basis for COVID-19 serological tests.

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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 10.1101/2020.12.04.407510 on bioRxiv

   Dec 4, 2020