Hetero-bivalent nanobodies provide broad-spectrum protection against SARS-CoV-2 variants of concern including Omicron

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1. Version published to 10.1038/s41422-022-00700-3

   Jul 29, 2022
2. 

   ScreenIT

   Mar 11, 2022

   SciScore for 10.1101/2022.03.08.483381: (What is this?)

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

   Table 1: Rigor

   Ethics	not detected.
   Sex as a biological variable	aRBD-2-5-Fc prophylactic protection in mice: Female K-18 hACE2 transgenic mice (The Jackson Laboratory) were administered with one dose of aRBD-2-5-Fc (10 mg/kg, i.p.) 24 hr before infected with 2 ×104 PFU of original SARS-CoV-2 (i.n.), and female A/J mice (The Jackson Laboratory) were administered with one dose of aRBD-2-5-Fc (1 mg/kg and 0.1 mg/kg, i.p.) 24 hr before infected with 1 ×105 PFU of mouse-adapted SARS-CoV-2 (intratracheally), body weight and death of the mice were monitored daily for 7 days post infection.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   …

   SciScore for 10.1101/2022.03.08.483381: (What is this?)

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

   Table 1: Rigor

   Ethics	not detected.
   Sex as a biological variable	aRBD-2-5-Fc prophylactic protection in mice: Female K-18 hACE2 transgenic mice (The Jackson Laboratory) were administered with one dose of aRBD-2-5-Fc (10 mg/kg, i.p.) 24 hr before infected with 2 ×104 PFU of original SARS-CoV-2 (i.n.), and female A/J mice (The Jackson Laboratory) were administered with one dose of aRBD-2-5-Fc (1 mg/kg and 0.1 mg/kg, i.p.) 24 hr before infected with 1 ×105 PFU of mouse-adapted SARS-CoV-2 (intratracheally), body weight and death of the mice were monitored daily for 7 days post infection.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   After four washes with PBST (PBS containing 0.1% tween-20), bound Nb-Fc were detected with a HRP-conjugated anti-IgG1 Fc antibody (Sino Biological).	anti-IgG1 suggested: None
   After 2days of infection, the cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton-100, blocked with DMEM containing 10% fetal bovine serum, and stained with a rabbit monoclonal antibody against SARS-CoV-2 NP (GeneTex, GTX635679) and an Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (Thermo Fisher Scientific)	SARS-CoV-2 NP suggested: None GTX635679 suggested: (GeneTex Cat# GTX635679, RRID:AB_2888553) anti-mouse suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   The recombinant vectors were transiently transfected into HEK293F cells with polyethyleneimine (Polyscience).	HEK293F suggested: RRID:CVCL_6642)
   Briefly, Vero E6 cells were seeded overnight in 24-well culture plates at 1.5 ×105 per well.	Vero E6 suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   aRBD-2-5-Fc prophylactic protection in mice: Female K-18 hACE2 transgenic mice (The Jackson Laboratory) were administered with one dose of aRBD-2-5-Fc (10 mg/kg, i.p.) 24 hr before infected with 2 ×104 PFU of original SARS-CoV-2 (i.n.), and female A/J mice (The Jackson Laboratory) were administered with one dose of aRBD-2-5-Fc (1 mg/kg and 0.1 mg/kg, i.p.) 24 hr before infected with 1 ×105 PFU of mouse-adapted SARS-CoV-2 (intratracheally), body weight and death of the mice were monitored daily for 7 days post infection.	K-18 hACE2 suggested: RRID:IMSR_GPT:T037657) A/J suggested: None
   Recombinant DNA
   Sentences	Resources
   Briefly, the coding sequences for SARS-CoV-2 RBD (amino acids 321 to 591) of original WT, Alpha (N501Y), Beta (K417N, E484K, N501Y), Gamma (K417T, E484K, N501Y), Delta (L452R, T478K), Delta plus (K417N, L452R, T478K), Kappa (L452R, E484Q) and Lambda (L452R, F490S) variant were cloned into pTT5 vector, which the C terminal contain a TEV cleavage site and a human IgG1 Fc.	pTT5 suggested: RRID:Addgene_52326)
   Software and Algorithms
   Sentences	Resources
   The data was analyzed using GraphPad Prism software.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)
   The total number of cells indicated by the nuclei staining and the infected cells indicated by the NP staining were quantified with the cellular analysis module of the Gen5 software (BioTek).	Gen5 suggested: (Gen5, RRID:SCR_017317)
   Subsequent models building and refinement were achieved using COOT and Phenix 62.	COOT suggested: (Coot, RRID:SCR_014222)
   All structural figures were prepared by PyMOL.	PyMOL suggested: (PyMOL, RRID:SCR_000305)

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

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3. 

   Version published to 10.1101/2022.03.08.483381v1 on bioRxiv

   Mar 8, 2022