ChAdOx1 nCoV-19 vaccine elicits monoclonal antibodies with cross-neutralizing activity against SARS-CoV-2 viral variants

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1. Version published to 10.1016/j.celrep.2022.110757

   May 1, 2022
2. 

   ScreenIT

   Oct 1, 2021

   SciScore for 10.1101/2021.09.27.461862: (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	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Fab/Fc ELISA: 96-well plates (Corning, 3690) were coated with goat anti-human Fc IgG antibody at 3 μg/mL overnight at 4°C.	anti-human Fc IgG suggested: None
   Secondary antibody, goat-anti-human-Fc-AP, was added at 1:1000 dilution in blocking buffer and incubated for 1 h at room temperature.	goat-anti-human-Fc-AP suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Biotinylated Spike or RBD were expressed in 1L of HEK293F cells (Invitrogen) at a density of 1.5 × 106 cells/mL.	HEK293F

   SciScore for 10.1101/2021.09.27.461862: (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	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Fab/Fc ELISA: 96-well plates (Corning, 3690) were coated with goat anti-human Fc IgG antibody at 3 μg/mL overnight at 4°C.	anti-human Fc IgG suggested: None
   Secondary antibody, goat-anti-human-Fc-AP, was added at 1:1000 dilution in blocking buffer and incubated for 1 h at room temperature.	goat-anti-human-Fc-AP suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Biotinylated Spike or RBD were expressed in 1L of HEK293F cells (Invitrogen) at a density of 1.5 × 106 cells/mL.	HEK293F suggested: None
   SARS-CoV-2 pseudotyped virus preparation: Pseudotyped HIV-1 virus incorporating either the SARS-Cov-2 Wuhan, B.1.1.7, P.1, B.1.351, B.1.617.2 full-length Spike were produced in a 10 cm dish seeded the day prior with 5×106 HEK293T/17 cells in 10 mL of complete Dulbecco’s Modified Eagle’s Medium (DMEM-C, 10% fetal bovine serum (FBS) and 1% Pen/Strep (100 IU/mL penicillin and 100 mg/mL streptomycin)).	HEK293T/17 suggested: ATCC Cat# CRL-11268, RRID:CVCL_1926)
   Gibson assembly products were directly transfected into HEK-293T cells and transformed under ampicillin selection.	HEK-293T suggested: CCLV Cat# CCLV-RIE 1018, RRID:CVCL_0063)
   IgG expression and purification: Ab heavy and light plasmids were co-transfected at a 1:1 ratio into HEK-293F cells (Thermofisher) using PEI Max (1 mg/mL, Polysciences, Inc.) at a 3:1 ratio (PEI Max:DNA).	HEK-293F suggested: RRID:CVCL_6642)
   HeLa-ACE2 and HeLa cells were washed once with PBS and detached using PBS containing 5mM EDTA.	HeLa-ACE2 suggested: None HeLa suggested: CLS Cat# 300194/p772_HeLa, RRID:CVCL_0030)
   Recombinant DNA
   Sentences	Resources
   Spike and RBD were cloned into a pHLsec vector containing Avi and 6xHis tags (Aricescu et al., 2006).	pHLsec suggested: None
   Software and Algorithms
   Sentences	Resources
   PBMCs were stained with live/dead (fixable Aqua Dead, Thermofisher), anti-CD3-APC/Cy7 (Biolegend), anti-CD8-APC-Cy7 (Biolegend), anti-CD14-BV510 (Biolegend), anti-CD19-PerCP-Cy5.5 (Biolegend), anti-IgM-PE (Biolegend), anti-IgD-Pacific Blue (Biolegend) and anti-IgG-PeCy7 (BD) and Spike-Alexa488 (Thermofisher Scientific, S32354) and Spike-APC (Thermofisher Scientific, S32362) or RBD-Alexa488 and RBD-APC.	Thermofisher suggested: (ThermoFisher; SL 8; Centrifuge, RRID:SCR_020809)
   D’Agostino & Pearson normality test, Kruskal-Wallis test with Dunn’s multiple comparisons post hoc test, Ordinary one-way ANOVA with Tukey’s multiple comparisons post hoc test and two-sided binomial tests) were performed using GraphPad Prism software.	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: We detected the following sentences addressing limitations in the study:

   Limitations of study: This study only examines mAbs isolated from one individual and therefore how representative these mAbs are of the humoral immune response against AZD1222 needs to be investigated further.

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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 10.1101/2021.09.27.461862 on bioRxiv

   Sep 28, 2021