Cross-reactivity of SARS-CoV structural protein antibodies against SARS-CoV-2

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1. 

   ScreenIT

   Mar 1, 2021

   SciScore for 10.1101/2020.07.30.229377: (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	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Transfected cells were incubated for 3 hours at RT with the following anti-SARS-CoV structural protein monoclonal or polyclonal antibodies at a 1:250 dilution:	anti-SARS-CoV structural protein suggested: None
   , mouse anti-SARS-CoV N monoclonal IgN 42c, rabbit anti-SARS-CoV S polyclonal sera (BEI Resources) and mouse anti-2xStrep-tag antibody (Sigma-Aldrich)	anti-SARS-CoV N suggested: None anti-2xStrep-tag suggested: None
   . Anti-mouse IgG AF555, anti-rabbit IgG …

   SciScore for 10.1101/2020.07.30.229377: (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	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Transfected cells were incubated for 3 hours at RT with the following anti-SARS-CoV structural protein monoclonal or polyclonal antibodies at a 1:250 dilution:	anti-SARS-CoV structural protein suggested: None
   , mouse anti-SARS-CoV N monoclonal IgN 42c, rabbit anti-SARS-CoV S polyclonal sera (BEI Resources) and mouse anti-2xStrep-tag antibody (Sigma-Aldrich)	anti-SARS-CoV N suggested: None anti-2xStrep-tag suggested: None
   . Anti-mouse IgG AF555, anti-rabbit IgG AF555, or anti-mouse IgM AF488 conjugated secondary antibodies were added at 1:500 dilution for 1 hour at RT (Invitrogen).	Anti-mouse IgG suggested: (R and D Systems Cat# AF555, RRID:AB_355438) anti-rabbit IgG suggested: (SouthernBiotech Cat# 4030-32, RRID:AB_2795940) anti-mouse IgM suggested: (SouthernBiotech Cat# 1021-30, RRID:AB_2794251)
   LzGreen SARS-COV-2 S pseudotyped lentivirus were mixed with 2-fold dilutions of the following monoclonal or polyclonal anti-SARS-CoV-2 S antibodies: mouse anti-SARS-CoV S monoclonal IgM 154c	anti-SARS-CoV-2 S suggested: None
   The following anti SARS-CoV-2 S monoclonal and polyclonal antibodies were serially diluted by 2-fold dilutions in blocking buffer: mouse anti-SARS-CoV S monoclonal IgM 154c	anti SARS-CoV-2 suggested: None anti-SARS-CoV S suggested: None
   Anti-mouse HRP, and anti-human-HRP secondary antibodies were used at 1:4000 concentration in blocking buffer, and were incubated 1 hour at RT. 50 μL of TMB HRP substrate (ThermoFisher Scientific) was added, and following incubation for 10 minutes at RT, 50μL of 2N H2SO4 was added as a stopping solution.	Anti-mouse HRP suggested: None anti-human-HRP secondary antibodies suggested: None anti-human-HRP suggested: None
   Resolved proteins were then transferred to a PVDF membrane, blocked in TBS with 2% BSA 0.1% Tween-20, then incubated with the following antibodies diluted to 1:500 in blocking buffer: mouse anti-SARS-CoV N monoclonal IgM 19c, mouse anti-SARS-CoV M monoclonal IgG1 283c, mouse anti-SARS-CoV E monoclonal IgM 472c, and mouse anti-2xStrep-tag antibody, and anti-His-HRP.	anti-SARS-CoV suggested: None anti-His-HRP suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Virus transduction capability was then tittered on 293T-Ace2 cells treated with 50μl of 5μg/ml polybrene (Sigma-Aldritch LLC).	293T-Ace2 suggested: RRID:CVCL_YZ65)
   His-tagged RBD bearing lentivirus was produced in HEK 293T cells and used to infect HEK 293-F suspension cells.	HEK 293T suggested: None
   Western blot: 293T cells were seeded in 10 cm dishes at a density of 3.5 million cells per dish.	293T suggested: None
   Software and Algorithms
   Sentences	Resources
   Maximum intensity z-projections were prepared in Fiji.	Fiji suggested: (Fiji, RRID:SCR_002285)

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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 10.1016/j.celrep.2021.108737

   Feb 1, 2021
3. 

   Version published to 10.1101/2020.07.30.229377v1 on bioRxiv

   Jul 30, 2020