Cornering an Ever-Evolving Coronavirus: TATX-03, a fully human synergistic multi-antibody cocktail targeting the SARS-CoV-2 Spike Protein with in vivo efficacy

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   ScreenIT

   Jul 26, 2021

   SciScore for 10.1101/2021.07.20.452858: (What is this?)

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

   Table 1: Rigor

   Ethics	Field Sample Permit: In vivo hamster challenge model of infection: All animal studies were performed at ViroClinics Xplore, Schaijk, The Netherlands and conducted according to European Union Directive 2010/63/EU and the standards of Dutch law for animal experimentation. Euthanasia Agents: On day 4 post-challenge all animals were euthanized by abdominal exsanguination under isoflurane anesthesia (3-5%).
   Sex as a biological variable	Briefly, groups of five specific pathogen free (SPF) male Syrian golden hamsters (Mesocricetus auratus) aged 9 to 10 weeks at the start of the experiment were randomly assigned to experimental groups.
   Randomization	Briefly, groups of five specific pathogen free (SPF) …

   SciScore for 10.1101/2021.07.20.452858: (What is this?)

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

   Table 1: Rigor

   Ethics	Field Sample Permit: In vivo hamster challenge model of infection: All animal studies were performed at ViroClinics Xplore, Schaijk, The Netherlands and conducted according to European Union Directive 2010/63/EU and the standards of Dutch law for animal experimentation. Euthanasia Agents: On day 4 post-challenge all animals were euthanized by abdominal exsanguination under isoflurane anesthesia (3-5%).
   Sex as a biological variable	Briefly, groups of five specific pathogen free (SPF) male Syrian golden hamsters (Mesocricetus auratus) aged 9 to 10 weeks at the start of the experiment were randomly assigned to experimental groups.
   Randomization	Briefly, groups of five specific pathogen free (SPF) male Syrian golden hamsters (Mesocricetus auratus) aged 9 to 10 weeks at the start of the experiment were randomly assigned to experimental groups.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   while full-length human IgG1 antibodies designated for in vitro neutralization and animal studies were produced in transiently transfected and/or stable CHO cells.	human IgG1 suggested: None
   Depletion panning with human ACE2- or CR3022-bound spike (where CR3022 represents an anti-SARS-CoV-1 antibody from the literature with cross-reactivity to SARS-CoV-223, 27) and irrelevant non-target proteins were performed to steer towards binders to specific epitopes and increase target specificity by reducing off-target reactivity, respectively.	anti-SARS-CoV-1 suggested: None SARS-CoV-223 suggested: None
   After washing with PBS-T, appropriate HRP-conjugated secondary antibody (anti-M13 and goat-anti-human-IgG for detection of phages and full size hIgG1, repsectively) was added and incubated for 60 minutes at RT.	anti-M13 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   In brief, HEK293-EBNA1 cells were transiently transfected with pUPE expression vectors containing recombinant protein encoding, codon-optimized synthetic genes.	HEK293-EBNA1 suggested: ATCC Cat# CRL-10852, RRID:CVCL_6974)
   Antibodies, expressed either as human IgG1-Fab and full-length human IgG1, destined for (initial) in vitro binding experiments were produced following transient transfection of HEK293 cells.	HEK293 suggested: CLS Cat# 300192/p777_HEK293, RRID:CVCL_0045)
   while full-length human IgG1 antibodies designated for in vitro neutralization and animal studies were produced in transiently transfected and/or stable CHO cells.	CHO suggested: CLS Cat# 603479/p746_CHO, RRID:CVCL_0213)
   To induce expression of spike trimers in a cell context, HEK293F cells were transiently transfected using FectoPRO per the manufacturer specification (PolyPlus Transfection, Illkirch, France) with SARS-CoV-2 surface glycoprotein expression vector.	HEK293F suggested: RRID:CVCL_6642)
   Briefly, the SARS-CoV-2 Spike protein was cloned into the pCAGGS expression vector system and transfected into HEK-293T cells.	HEK-293T suggested: CCLV Cat# CCLV-RIE 1018, RRID:CVCL_0063)
   Vero E6 monolayers are washed and incubated for 5 or 6 days at 37 °C.	Vero E6 suggested: None
   Recombinant DNA
   Sentences	Resources
   The fragments were ligated into the pHENIX-His8-VSV vector and transformed into Escherichia coli TG1 cells.	pHENIX-His8-VSV suggested: None
   Briefly, the SARS-CoV-2 Spike protein was cloned into the pCAGGS expression vector system and transfected into HEK-293T cells.	pCAGGS suggested: RRID:Addgene_18926)
   Software and Algorithms
   Sentences	Resources
   Library panning: The phage display libraries employed here were previously generated by ImmunoPrecise Antibodies (Naïve Human Library #0899	Human Library suggested: None
   Heat maps were curated manually in Excel by merging the results from different experiments.	Excel suggested: None
   EC50 values were calculated in GraphPad Prism.	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 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.
   • No funding statement was detected.
   • 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 10.1101/2021.07.20.452858v1 on bioRxiv

   Jul 21, 2021