Structural basis of a two-antibody cocktail exhibiting highly potent and broadly neutralizing activities against SARS-CoV-2 variants including diverse Omicron sublineages

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1. Version published to 10.1038/s41421-022-00449-4

   Sep 8, 2022
2. 

   Version published to 10.1101/2022.05.27.493682v3 on bioRxiv

   Jun 1, 2022
3. 

   Version published to 10.1101/2022.05.27.493682v2 on bioRxiv

   May 30, 2022
4. 

   ScreenIT

   May 29, 2022

   SciScore for 10.1101/2022.05.27.493682: (What is this?)

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

   Table 1: Rigor

   Ethics	Euthanasia Agents: At the end point of the experiment, all remaining animals in the monoclonal antibody-administered group received an overdose of isoflurane and were humanely euthanized. IACUC: Ethics statement: This study was approved by the Experimental Animal Welfare and Ethical Review Board of Wuhan Institute of Biological Products Co.
   Sex as a biological variable	Male K18-hACE2 mice (6–8 weeks old, purchased from GemPharmatech Co., Ltd. Company.) were randomly distributed into groups (n = 3–6 mice per group).
   Randomization	Male K18-hACE2 mice (6–8 weeks old, purchased from GemPharmatech Co., Ltd. Company.) were randomly distributed into groups (n = 3–6 mice per group).
   Blinding	not detected.
   Po…

   SciScore for 10.1101/2022.05.27.493682: (What is this?)

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

   Table 1: Rigor

   Ethics	Euthanasia Agents: At the end point of the experiment, all remaining animals in the monoclonal antibody-administered group received an overdose of isoflurane and were humanely euthanized. IACUC: Ethics statement: This study was approved by the Experimental Animal Welfare and Ethical Review Board of Wuhan Institute of Biological Products Co.
   Sex as a biological variable	Male K18-hACE2 mice (6–8 weeks old, purchased from GemPharmatech Co., Ltd. Company.) were randomly distributed into groups (n = 3–6 mice per group).
   Randomization	Male K18-hACE2 mice (6–8 weeks old, purchased from GemPharmatech Co., Ltd. Company.) were randomly distributed into groups (n = 3–6 mice per group).
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Then cells were stained with anti-mouse IgG Taxes red conjugated antibody and anti-human IgG FITC-conjugated antibody (Sigma, USA) for another 30 min then analyzed by FACS Aria II (BD, USA).	anti-mouse IgG suggested: None anti-human IgG suggested: None
   Antibody Binding Kinetics Measured by SPR: The binding kinetics of mAbs to SARS-CoV-2 Delta-RBD or Omicron-RBD monomer were analyzed using SPR (Biacore 8K; GE Healthcare).	SARS-CoV-2 suggested: None Delta-RBD suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Cells, Viruses and Proteins: Cell lines (HEK293T and Vero E6 cells) were initially acquired from the American Type Culture Collection (ATCC; USA).	Vero E6 suggested: RRID:CVCL_XD71)
   HEK293T-hACE2-cells were generated via the overexpression of the human ACE2 receptor in HEK293T cells and were used in the neutralization assays of pseudoviruses.	HEK293T-hACE2-cells suggested: None
   Then mixtures were added to 2.5 × 105 HEK293T cells expressing ACE2 and incubated at 4 °C for another hour.	HEK293T suggested: RRID:CVCL_HA71)
   HEK293-hACE2 cells (2.5 × 104 cells/100μL per well) were then added into the mixture and incubated at 37 °C in a humidified atmosphere with 5% CO2 for 23 h to 25 h.	HEK293-hACE2 suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Male K18-hACE2 mice (6–8 weeks old, purchased from GemPharmatech Co., Ltd. Company.) were randomly distributed into groups (n = 3–6 mice per group).	K18-hACE2 suggested: RRID:IMSR_GPT:T037657)
   Recombinant DNA
   Sentences	Resources
   The SARS-CoV-2 Spike ectodomain (1-1208) with a C-terminal Strep tag for purification and a foldon tag for trimerization was inserted into the pFastBac-Dual vector (Invitrogen) and was expressed using Bac-to-Bac baculovirus system (Invitrogen).	pFastBac-Dual suggested: RRID:Addgene_137166)
   The cDNA encoding SARS-CoV-2 Omicron Spike was synthesized (GenBank ID: ULC25168.1) and cloned into the pCAG vector.	pCAG suggested: RRID:Addgene_74288)
   Software and Algorithms
   Sentences	Resources
   All of these data were analyzed using Flow Jo.	Flow Jo suggested: (FlowJo, RRID:SCR_008520)
   All statistical analysis was performed using GraphPad Prism 8.	GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)
   Coot v.	Coot suggested: (Coot, RRID:SCR_014222)
   Figures were generated using PyMOL 2.0.779	PyMOL suggested: (PyMOL, RRID:SCR_000305)

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   Results from OddPub: Thank you for sharing your data.

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

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   Read the original source
5. 

   Version published to 10.1101/2022.05.27.493682v1 on bioRxiv

   May 27, 2022