Evaluation of a commercial ELISA as alternative to plaque reduction neutralization test to detect neutralizing antibodies against SARS-CoV-2

Article activity feed

1. Version published to 10.1038/s41598-022-07597-3

   Mar 3, 2022
2. 

   ScreenIT

   Oct 17, 2021

   SciScore for 10.1101/2021.10.12.21264881: (What is this?)

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

   Table 1: Rigor

   Ethics	not detected.
   Sex as a biological variable	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Neutralizing antibodies are considered to block the binding between RBD and ACE2, hence increasing the amount of free RBD that can bind to the plate.	ACE2 suggested: None
   Next, the plate was washed 4x with the supplied wash buffer and 100 μL of Spike-RBD antibodies conjugated to HRP were added per well.	Spike-RBD suggested: None
   Binding antibody ELISA tests: The Euroimmun SARS-CoV-2 IgG antibody ELISA (“S1 IgG”), Euroimmun SARS-CoV-2-NCP IgG ELISA (“NCP IgG”, both Euroimmun AG, Lübeck, …

   SciScore for 10.1101/2021.10.12.21264881: (What is this?)

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

   Table 1: Rigor

   Ethics	not detected.
   Sex as a biological variable	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Neutralizing antibodies are considered to block the binding between RBD and ACE2, hence increasing the amount of free RBD that can bind to the plate.	ACE2 suggested: None
   Next, the plate was washed 4x with the supplied wash buffer and 100 μL of Spike-RBD antibodies conjugated to HRP were added per well.	Spike-RBD suggested: None
   Binding antibody ELISA tests: The Euroimmun SARS-CoV-2 IgG antibody ELISA (“S1 IgG”), Euroimmun SARS-CoV-2-NCP IgG ELISA (“NCP IgG”, both Euroimmun AG, Lübeck, Germany) and the Wantai SARS-CoV-2 Ab ELISA (Beijing Wantai Biological Pharmacy Enterprise; Beijing, China) were carried out according to the manufacturer’s instructions, with the exception that 50 μl of serum were used in the Wantai Ab ELISA.	The Euroimmun SARS-CoV-2 IgG antibody ELISA suggested: None SARS-CoV-2 IgG suggested: None “S1 IgG” suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Next, 100 μL of diluted serum–virus mix were added to wells containing 2 × 104 Vero E6 cells per well (#85020206, European Collection of Authenticated Cell Cultures (ECACC), Porton Down, UK) in a 96-well plate.	Vero E6 suggested: None

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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 some technical limitations concerning the use of the GenScript sVNT for high-throughput sample analysis. We have identified pipetting speed as one of the main factors involved in the generation of false positive results, as the strength of the signal is strongly correlated with incubation time. If pipetting occurs slowly (e.g. when using a single-channel pipette), HRP-coupled RBD will have more time to bind to the coated ACE receptor, and will thus create a stronger OD signal, in negative samples at the beginning of the plate compared to negative samples at the end of the plate. In our laboratory, where the GenScript sVNT is performed manually, we were able to address this “OD drift” over the plate by using a multichannel pipette and by processing only half plates at a time. However, for full automation this time-dependent drift can be problematic as many pipetting robots use a single-tip dispenser and, hence, robots often cannot adhere to the 2-min pipetting time per plate as recommended by the manufacturer. Nonetheless, the throughput achievable by using the GenScript sVNT remains much higher than that of a cell-culture-based PRNT, even if only half plates are processed. However, it impairs the large-scale testing of vaccinated individuals on a routine basis. Another parameter that remains to be evaluated when using the GenScript sVNT is the influence of emerging variants with mutations in the RBD, e.g. B1.1.7, B.1.351 and P1. A possible solution would be the deve...

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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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   Results from scite Reference Check: We found no unreliable references.

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3. 

   Version published to 10.1101/2021.10.12.21264881 on medRxiv

   Oct 14, 2021