Dimerization of SARS-CoV-2 nucleocapsid protein affects sensitivity of ELISA based diagnostics of COVID-19

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1. Version published to 10.1016/j.ijbiomac.2022.01.094

   Mar 1, 2022
2. 

   Version published to 10.1101/2021.05.23.445305v2 on bioRxiv

   May 27, 2021
3. 

   ScreenIT

   May 26, 2021

   SciScore for 10.1101/2021.05.23.445305: (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.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The membrane was then washed thrice with 1X-TBST and incubated with anti-SARS-CoV-2 N protein antibodies (catalogue No. ab272852, Abcam, 1:1000) in TBST with BSA (2%, w/v).	anti-SARS-CoV-2 N protein suggested: None
   Immunoblot was then incubated with HRP conjugated-goat anti-human IgG secondary antibodies (Millipore, AP309P) in a dilution of 1:10,000 for 1 h.	anti-human IgG suggested: (Millipore Cat# AP309P, RRID:AB_11212373)
   The anti-SARS-CoV-2 N protein antibodies (Abcam) were diluted in 1X TBST and 100 μL of the diluted …

   SciScore for 10.1101/2021.05.23.445305: (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.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The membrane was then washed thrice with 1X-TBST and incubated with anti-SARS-CoV-2 N protein antibodies (catalogue No. ab272852, Abcam, 1:1000) in TBST with BSA (2%, w/v).	anti-SARS-CoV-2 N protein suggested: None
   Immunoblot was then incubated with HRP conjugated-goat anti-human IgG secondary antibodies (Millipore, AP309P) in a dilution of 1:10,000 for 1 h.	anti-human IgG suggested: (Millipore Cat# AP309P, RRID:AB_11212373)
   The anti-SARS-CoV-2 N protein antibodies (Abcam) were diluted in 1X TBST and 100 μL of the diluted antibodies were allowed to interact with the coated N protein in the ELISA wells at 37°C for 1 h.	The anti-SARS-CoV-2 N protein antibodies ( Abcam ) suggested: None anti-SARS-CoV-2 N protein antibodies ( Abcam ) suggested: None
   Wells were then washed with 200 μL of 1X TBST buffer three times followed by incubation with 100 μL of goat IgG-HRP antibody (Thermo Fisher Scientific) prepared in 1X TBST buffer.	goat IgG-HRP suggested: None
   Recombinant DNA
   Sentences	Resources
   The expression construct of SARS-CoV-2 N protein was commercially procured through Addgene in a pGBW-m4046785 vector with N protein gene insert of 1253 bp under the control of T7 promoter.	pGBW-m4046785 suggested: RRID:Addgene_145684)
   Software and Algorithms
   Sentences	Resources
   MS spectrum was analysed with Agilent MassHunter Qualitative analysis software (B.07.00).	Agilent MassHunter suggested: (Agilent MassHunter Qualitative Analysis, RRID:SCR_019081)
   Pymol tool was used to superimpose the structure of 6WZO and modelled monomeric structure to build its dimeric form.	Pymol suggested: (PyMOL, RRID:SCR_000305)
   Later, a similar analysis was performed with the DiscoTope server.	DiscoTope suggested: (DiscoTope, RRID:SCR_018530)

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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.
   • No funding statement was detected.
   • No protocol registration statement was detected.

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

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4. 

   Version published to 10.1101/2021.05.23.445305v1 on bioRxiv

   May 24, 2021