Development of a rapid point-of-care test that measures neutralizing antibodies to SARS-CoV-2

Article activity feed

1. Version published to 10.1016/j.jcv.2021.105024

   Dec 1, 2021
2. 

   ScreenIT

   Dec 16, 2020

   SciScore for 10.1101/2020.12.15.20248264: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: Methods Human Subjects and Samples Peripheral blood, serum and plasma were collected for this study under an Arizona State University institutional review board (IRB) approved protocol #0601000548 and Mayo Clinic IRB protocol #20-004544.
   Randomization	not detected.
   Blinding	( De-identified serum samples were sent from Mayo Clinic Rochester to our laboratories at Mayo Clinic Arizona and run 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
   A separate set of 38 samples was run concurrently in the SARS-CoV-2 authentic microneutralizing …

   SciScore for 10.1101/2020.12.15.20248264: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: Methods Human Subjects and Samples Peripheral blood, serum and plasma were collected for this study under an Arizona State University institutional review board (IRB) approved protocol #0601000548 and Mayo Clinic IRB protocol #20-004544.
   Randomization	not detected.
   Blinding	( De-identified serum samples were sent from Mayo Clinic Rochester to our laboratories at Mayo Clinic Arizona and run 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
   A separate set of 38 samples was run concurrently in the SARS-CoV-2 authentic microneutralizing antibody assay, the LFA, and the Ortho Vitros Anti-SARS-CoV-2 IgG assay.	Anti-SARS-CoV-2 IgG suggested: None
   For procedural control purposes, the LFA also contains a control mouse antibody conjugated to red gold nanospheres and corresponding anti-mouse control line.	anti-mouse control line. suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   The pseudotyped virus is called VSV- SARS-CoV-2-S-∆19CT and was produced by BHK-21 cells.	BHK-21 suggested: ATCC Cat# CRL-6282, RRID:CVCL_1914)
   Two different Vero cells encoding split product luciferase (DSP1 and DSP2) were cultured at a 1:1 ratio as a monolayer.	Vero suggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)
   Software and Algorithms
   Sentences	Resources
   Correlation analysis was conducted using IBM SPSS (Armonk, NY).	SPSS suggested: (SPSS, RRID:SCR_002865)
   Regression analysis was performed using Microsoft Excel (Redmond, WA); Bland-Altman plots were visualized using IBM SPSS.	Microsoft Excel suggested: (Microsoft Excel, RRID:SCR_016137)
   ROC analysis was conducted using R language in the RStudio environment (version 3.6.2; RStudio PBC, Boston, MA).	RStudio suggested: (RStudio, RRID:SCR_000432)

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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:

   Limitations of the LFA are that it uses only the RBD portion of SARS-CoV-2 spike protein. Although the vast majority of reports indicate that the principle neutralizing domain is RBD portion of spike protein, mAbs have been reported that neutralize SARS-CoV-2 by binding to the N-terminal domain of spike protein(36, 37). Also, since the spike protein assumes several conformations during viral binding and entry(38), neutralizing epitopes exist on the quaternary structure of spike(37). Although RBDs on the nanoparticles may associate, it is unlikely they assume a native quaternary conformation. Other limitations are the binary nature of the data analysis (neutralizing and non- neutralizing) of a continuous assay. Clearly, line densities demonstrate moderate levels of neutralization. Since blood draws and subsequent assays are a “snapshot in time” of neutralizing antibody activity, levels were undoubtedly increasing in some patients and decreasing in others. LFAs are generally inexpensive and highly portable compared to other laboratory-based tests, so neutralizing antibody levels could be measured using the LFA to longitudinally assess protective neutralizing antibody immunity. Another limitation is that the LFA does not differentiate high affinity anti-RBD NAbs from an abundance of lower affinity anti- RBD NAbs. In related experiments, we have observed patient sera that bind strongly to RBD, but do not demonstrate neutralizing activity (data not shown). This test may prove very...

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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 10.1101/2020.12.15.20248264 on medRxiv

   Dec 16, 2020