A key linear epitope for a potent neutralizing antibody to SARS-CoV-2 S-RBD

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   ScreenIT

   Sep 12, 2020

   SciScore for 10.1101/2020.09.11.292631: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	Consent: The original clinical studies to obtain blood samples after written informed consent were previously described6 and had been approved by the Ethics Board of ChongQing Medical University. IRB: The original clinical studies to obtain blood samples after written informed consent were previously described6 and had been approved by the Ethics Board of ChongQing Medical University.
   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
   The antibodies were isolated using flow sorting for isolation and cloning of single …

   SciScore for 10.1101/2020.09.11.292631: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	Consent: The original clinical studies to obtain blood samples after written informed consent were previously described6 and had been approved by the Ethics Board of ChongQing Medical University. IRB: The original clinical studies to obtain blood samples after written informed consent were previously described6 and had been approved by the Ethics Board of ChongQing Medical University.
   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
   The antibodies were isolated using flow sorting for isolation and cloning of single antigen-specific B cells and the antibody variable genes encoding monoclonal antibodies6.	antibodies6 suggested: None
   Recombinant antibody production and purification: For the construction of antibody expression Vectors, VH and VL 2nd PCR products were inserted separately into the linearized plasmids (pcDNA3.4) that encode constant regions of the heavy chains and light chains via a homologous recombination kit (Vazyme, C112).	C112 suggested: None
   Sequence analysis of antigen-specific mAb: IMGT/V-QUEST (http://www.imgt.org/IMGT_vquest/vquest) and IgBLAST (https://www.ncbi.nlm.nih.gov/igblast/), MIXCR (https://mixcr.readthedocs.io/en/master/) and VDJtools (https://vdjtools-doc.readthedocs.io/en/master/overlap.html) tools were used to do the variable region analysis and annotation for each antibody clone.	antigen-specific mAb: IMGT/V-QUEST (http://www.imgt.org/IMGT_vquest/vquest) suggested: None
   A CM5 chip (GE Healthcare) was linked with anti-human IgG-Fc antibody to capture about 9000 response units of the NAbs.	anti-human IgG-Fc suggested: None
   After 30 min incubation at 37 °C, the plates were washed 5 times and incubated with goat anti-human IgG (H+L) antibody conjugated with ALP (Thermo Fisher, a18808, 1:5000) for 30 min at 37 °C.	anti-human IgG suggested: (Thermo Fisher Scientific Cat# A18808, RRID:AB_2535585) a18808 suggested: (Thermo Fisher Scientific Cat# A18808, RRID:AB_2535585)
   Reacted mAbs were detected using ALP-conjugated Goat F(ab’)2 Anti-Human (IgG (Fab’)2) secondary antibody (Abcam, ab98532, 1:2000) for 30 min at RT, followed with quantification detection.	F(ab’)2 Anti-Human (IgG suggested: None
   The ELISA plates were washed 4 times by Blocking Buffer and 50 μL Goat F(ab’)2 Anti-Human (IgG (Fab’)2) secondary antibody conjugated with ALP (Abcam, ab98532, 1:2000) was incubated with the plate at RT for 30 min.	Anti-Human suggested: None
   The next days, the membranes were washed with TBST and incubated with HRP-conjugated Goat-anti-human Fc antibody (Abcam, ab99759, 1:10000) for 1 h at RT.	ab99759 suggested: (Abcam Cat# ab99759, RRID:AB_10673762)
   Experimental Models: Cell Lines
   Sentences	Resources
   HEK293T cells were grown to 80% confluency before transfection with VSV-G pseudotyped ΔG-luciferase, pWPXL and pSPAX2.	HEK293T suggested: None
   After 72 hrs, the luciferase activities of infected HEK293T/ACE2 cells were detected by the Bright-Luciferase Reporter Assay System (Promega, E2650).	HEK293T/ACE2 suggested: None
   After the incubation, the mixtures were then transferred into 96-well plates, which were seeded with Vero E6 cells.	Vero E6 suggested: RRID:CVCL_XD71)
   Software and Algorithms
   Sentences	Resources
   Sequence analysis of antigen-specific mAb: IMGT/V-QUEST (http://www.imgt.org/IMGT_vquest/vquest) and IgBLAST (https://www.ncbi.nlm.nih.gov/igblast/), MIXCR (https://mixcr.readthedocs.io/en/master/) and VDJtools (https://vdjtools-doc.readthedocs.io/en/master/overlap.html) tools were used to do the variable region analysis and annotation for each antibody clone.	IgBLAST suggested: (IgBLAST, RRID:SCR_002873)
   The half-maximal inhibitory concentrations (IC50) of the evaluated mAbs were tested by the Varioskan LUX Microplate Spectrophotometer (Thermo Fisher), and calculated by a four-parameter logistic regression using GraphPad Prism 8.0.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)

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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.
   • 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 10.1101/2020.09.11.292631v1 on bioRxiv

   Sep 11, 2020