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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Abstract
The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has created an ongoing global human health crisis and will likely become endemic, requiring novel sustainable therapeutic strategies. We report on the discovery of a fully human multi-antibody cocktail (TATX-03) targeting diversified non-overlapping epitopes on the SARS-CoV-2 spike protein that suppressed replication-competent viral titers to undetectable levels in the lungs of SARS-CoV-2 challenged hamsters upon both prophylactic and therapeutic administration. While monotherapy with two of the individual cocktail components also showed clear in vivo protection, neither recapitulated the efficacy of TATX-03. This synergistic effect was further supported by examining in vivo efficacy of these individual antibodies and corresponding combination therapy at a lower dose. Furthermore, in vitro screenings using VSV-particles pseudo-typed with spike proteins representing the SARS-CoV-2 variants of concern Alpha, Beta, and Delta showed that TATX-03 maintained its neutralization potency. These results merit further development of TATX-03 as a potential therapy for SARS-CoV-2 infection with resistance to mutagenic escape.
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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 IgG1suggested: NoneDepletion 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-1suggested: NoneSARS-CoV-223suggested: NoneAfter 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-M13suggested: NoneExperimental 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-EBNA1suggested: 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. HEK293suggested: 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. CHOsuggested: 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. HEK293Fsuggested: 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-293Tsuggested: CCLV Cat# CCLV-RIE 1018, RRID:CVCL_0063)Vero E6 monolayers are washed and incubated for 5 or 6 days at 37 °C. Vero E6suggested: NoneRecombinant DNA Sentences Resources The fragments were ligated into the pHENIX-His8-VSV vector and transformed into Escherichia coli TG1 cells. pHENIX-His8-VSVsuggested: NoneBriefly, the SARS-CoV-2 Spike protein was cloned into the pCAGGS expression vector system and transfected into HEK-293T cells. pCAGGSsuggested: 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 Librarysuggested: NoneHeat maps were curated manually in Excel by merging the results from different experiments. Excelsuggested: NoneEC50 values were calculated in GraphPad Prism. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)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).
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.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
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.
Results from scite Reference Check: We found no unreliable references.
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