Engineering SARS-CoV-2 cocktail antibodies into a bispecific format improves neutralizing potency and breadth

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   ScreenIT

   Feb 4, 2022

   SciScore for 10.1101/2022.02.01.478504: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: Neutralization assays: All SARS-CoV-2 manipulations were conducted at the Biosafety Level-3 facility with the approval from the Institutional Biosafety Committee at the University of Texas Medical Branch.
   Sex as a biological variable	Female BALB/c mice aged 10-12 weeks (n = 10) were infected intranasally (IN) with 104 PFU of mouse-adapted SARS-CoV-2 CMA4 strain50 or the Beta and Gamma variants23 in 50 μl of PBS.
   Randomization	Seven-week-old female BALB/c (Jackson lab, USA) were randomly divided into three groups (5 mice/group) and were injected by i.p with 10 mg/kg of antibody.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   S…

   SciScore for 10.1101/2022.02.01.478504: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: Neutralization assays: All SARS-CoV-2 manipulations were conducted at the Biosafety Level-3 facility with the approval from the Institutional Biosafety Committee at the University of Texas Medical Branch.
   Sex as a biological variable	Female BALB/c mice aged 10-12 weeks (n = 10) were infected intranasally (IN) with 104 PFU of mouse-adapted SARS-CoV-2 CMA4 strain50 or the Beta and Gamma variants23 in 50 μl of PBS.
   Randomization	Seven-week-old female BALB/c (Jackson lab, USA) were randomly divided into three groups (5 mice/group) and were injected by i.p with 10 mg/kg of antibody.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The blocking percentages at each concentrations were calculated as: (normalized ACE2 response of isotype antibody-normalized ACE2 response of tested antibody)/ normalized ACE2 response of isotype antibody *100.	antibody-normalized ACE2 suggested: None
   The anti-human IgG Fab2 HRP-conjugated antibody was diluted 1:5000 and added at a volume of 100 μl per well for incubation at 37°C for 1h.	anti-human IgG suggested: None
   The HRP-conjugated goat anti-human IgG-F(ab’)2 was used as the secondary antibody and incubated at room temperature for 1 h.	anti-human IgG-F(ab’)2 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Vero (ATCC® CCL-81) and Vero E6 cells (ATCC, CRL-1586) were maintained in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% FBS.	Vero suggested: ATCC Cat# CRL-1586, RRID:CVCL_0574)
   After 1 h incubation at 37 °C, the antibody-virus mixtures were inoculated onto a monolayer of Vero E6 cells pre-seeded on 6-well plates on the previous day.	Vero E6 suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   For mouse study with the Delta variant, the 8-10-week-old female K18-hACE2 mice were ordered from The Jackson Laboratory.	K18-hACE2 suggested: RRID:IMSR_GPT:T037657)
   Seven-week-old female BALB/c (Jackson lab, USA) were randomly divided into three groups (5 mice/group) and were injected by i.p with 10 mg/kg of antibody.	BALB/c suggested: None
   Software and Algorithms
   Sentences	Resources
   ForteBio’s data analysis software was used to export data, and the binding profile was processed by GraphPad prism 8 Software.	GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)
   The initial model for the complex between 14-H-06 with four RBD molecules (one RBD bound for each of the four paratopes of IgG-scFv bsAb 14-H-06) was subjected to MD simulations using NAMD 2.1246.	NAMD suggested: (NAMD, RRID:SCR_014894)
   Structure analysis and image production were made using PyMOL (https://pymol.org, Schrödinger Inc.) and COOT49.	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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   Version published to 10.1101/2022.02.01.478504 on bioRxiv

   Feb 1, 2022