Development of SARS-CoV-2 Nucleocapsid Specific Monoclonal Antibodies

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   ScreenIT

   Sep 9, 2020

   SciScore for 10.1101/2020.09.03.280370: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Two 6-week old female BALB/c mice (Jackson Laboratories) were primed with antigen emulsified in an equal volume of complete Freund’s adjuvant (Millipore-Sigma).
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   After three more washes the plates were incubated with HRP conjugated goat anti-mouse polyclonal antibodies (Abcam ab97023) diluted at 1:10,000 in 1X PBS for 1 hour at room temperature, shaking.	anti-mouse polyclonal antibodies suggested: None
   Supernatant from parents 17, 21, 22, 57, and 67 along with rabbit …

   SciScore for 10.1101/2020.09.03.280370: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Two 6-week old female BALB/c mice (Jackson Laboratories) were primed with antigen emulsified in an equal volume of complete Freund’s adjuvant (Millipore-Sigma).
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   After three more washes the plates were incubated with HRP conjugated goat anti-mouse polyclonal antibodies (Abcam ab97023) diluted at 1:10,000 in 1X PBS for 1 hour at room temperature, shaking.	anti-mouse polyclonal antibodies suggested: None
   Supernatant from parents 17, 21, 22, 57, and 67 along with rabbit anti-N protein polyclonal control antibody were then incubated for 1 hour shaking at room temperature and then washed three times with 1X PBS + 0.1% Tween 20.	anti-N suggested: None
   FITC-conjugated goat anti-mouse secondary antibodies or Alexa 488-conjugated goat anti-rabbit secondary antibodies at a dilution of 1:2000 in 1X PBS were added to the appropriate wells, with all wells receiving Hoescht staining at 1:4000.	anti-mouse suggested: None anti-rabbit suggested: None
   Blots were washed three time with blocking solution for five minutes each before being incubated for 1 hour at room temperature with goat anti-mouse HRP-conjugated secondary antibodies diluted in blocking solution.	anti-mouse HRP-conjugated secondary suggested: None
   Sequencing of anti-N protein monoclonal antibody genes: RNA from each hybridomas 17, 21, 22, 57, and 67 was isolated and stored at −80°C following Trizol (Invitrogen) and phenol-chloroform extraction.	anti-N protein suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Stocks of virus were grown in Vero-E6 cells in DMEM (Gibco) supplemented with 10% fetal bovine serum (Atlas Biologicals) and 25mM HEPES (pH 7.5) in BSL-3+ containment at the Colorado State University Regional Containment Biocontainment Laboratory	Vero-E6 suggested: None
   Vero cells were infected at 0.1 MOI with SARS-CoV-2 in BSL-3 and allowed to incubate for 48 hours before cells were trypsinized, spun down, and resuspended in RIPA buffer.	Vero suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Two 6-week old female BALB/c mice (Jackson Laboratories) were primed with antigen emulsified in an equal volume of complete Freund’s adjuvant (Millipore-Sigma).	BALB/c suggested: RRID:IMSR_ORNL:BALB/cRl)
   Software and Algorithms
   Sentences	Resources
   Absorbances were corrected against the PBS negative control and organized by absorbance on Microsoft Excel.	Microsoft Excel suggested: (Microsoft Excel, RRID:SCR_016137)
   Five kappa and heavy chain clones were sequenced for each hybridoma by Genewiz.	Genewiz suggested: (GENEWIZ, RRID:SCR_003177)
   Clone sequences were analyzed by IgBLAST (30) for antibody framework regions (FMR) and complementary-determining regions (CDR).	IgBLAST suggested: (IgBLAST, RRID:SCR_002873)

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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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2. 

   Version published to 10.1101/2020.09.03.280370 on bioRxiv

   Sep 3, 2020