ZRC3308 monoclonal antibody cocktail shows protective efficacy in Syrian hamsters against SARS-CoV-2 infection

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   ScreenIT

   Sep 21, 2021

   SciScore for 10.1101/2021.09.16.460724: (What is this?)

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

   Table 1: Rigor

   Ethics

   IRB: Ethics statement: The study was approved by Institute Animal Ethics committee. IACUC: The study was performed according to the guidelines of Committee for the Purpose of Control and Supervision of Experiments on Animals, Government of India. mAb (ZRC3308-A7 and ZRC3308-B10) generation: The codon optimized, light and heavy chain genes of ZRC3308-A7 (CAS RN: 2640223-84-1) and ZRC3308-B10 (CAS RN: 2640224-48-0) were cloned in dual assembly eukaryotic expression vector, DGV-GS, with Glutamine synthase selection marker (DGV-GS). Euthanasia Agents: Four animals each of the prophylactic and therapeutic study group were sacrificed by overdose of isoflurane anesthesia on 3, 5 and 7 DPI and …

   SciScore for 10.1101/2021.09.16.460724: (What is this?)

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

   Table 1: Rigor

   Ethics	IRB: Ethics statement: The study was approved by Institute Animal Ethics committee. IACUC: The study was performed according to the guidelines of Committee for the Purpose of Control and Supervision of Experiments on Animals, Government of India. mAb (ZRC3308-A7 and ZRC3308-B10) generation: The codon optimized, light and heavy chain genes of ZRC3308-A7 (CAS RN: 2640223-84-1) and ZRC3308-B10 (CAS RN: 2640224-48-0) were cloned in dual assembly eukaryotic expression vector, DGV-GS, with Glutamine synthase selection marker (DGV-GS). Euthanasia Agents: Four animals each of the prophylactic and therapeutic study group were sacrificed by overdose of isoflurane anesthesia on 3, 5 and 7 DPI and necropsy was performed to collect nasal wash, nasal turbinates, lungs and blood samples.
   Sex as a biological variable	Pharmacokinetic study in hamsters: A total of 18 female hamsters, aged 11-12 weeks were divided into four groups according to the dose of the cocktail i.e., 50 mg/kg (25 mg/kg of ZRC3308-A7 + 25 mg/kg of ZRC-3308-B10), 5 mg/kg (2.5 mg/kg of ZRC3308-A7 + 2.5 mg/kg of ZRC3308-B10), 1.0 mg/kg (0.5 mg/kg of ZRC3308-A7 + 0.5 mg/kg of ZRC3308-B10), and placebo consisting of 5 animals each except for the placebo which consisted of only 3 animals.
   Randomization	not detected.
   Blinding	The lesions were blindly scored based on the vascular changes, bronchial lesions, alveolar pathological changes like septal thickening, pneumocyte hyperplasia, consolidation, edematous changes and inflammatory cell infiltration.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Detection was accomplished using 1/1000 diluted peroxidase conjugated sheep anti-C1q polyclonal antibody (Abcam, UK).	anti-C1q suggested: None
   ELISA based detection of mAb levels in serum: Two separate ELISA methods were used to detect ZRC3308-A7 and ZRC3308-B10 antibodies.	ZRC3308-B10 suggested: None
   Detection was accomplished using 1/100,000 diluted peroxidase conjugated goat anti-human lambda light chain secondary antibody for ZRC3308-A7 (Novus Biologics, USA) and 1/135,000 diluted Goat anti-Human kappa light chain secondary antibody for ZRC3308-B10 (Novus Biologics, USA) to specifically detect the two antibodies.	anti-human lambda light chain suggested: None anti-Human kappa light chain suggested: None
   The groups were high (50mg/kg), medium (5 mg/kg), low (1mg/kg) dose of mAb cocktail, an IgG1 isotype antibody (Trastuzumab @ 50 mg/kg) control and a placebo control.	an IgG1 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   HEK 293 ACE2 expressing cells (1.0×104 per well) were seeded in a flat tissue culture plate.	HEK 293 suggested: None
   Vero E6 cells (1.0×106 per well) were seeded to 24-well plates in maintenance medium for 24 hrs at 37°C in a 5% CO2 incubator.	Vero E6 suggested: None
   Software and Algorithms
   Sentences	Resources
   GraphPad Prism 8, Inc., San Diego, CA, USA).	GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)
   Statistical analysis: Graphpad Prism version 8.4.3 software was used for data analysis of the experimental challenge study.	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:

   However, the 6-hour therapeutic group had limitation of small sample size. The interim results of the phase 1-3 clinical trials (ClinicalTrials.gov number: NCT04425629) of the mAb REGN-COV2 in patients with early infection showed a reduced SARS-CoV-2 RNA level in nasopharyngeal swab samples following mAb administration19. Similar results were reported in case of bamlanivimab (LY-CoV555) treatment in patients with a median of 4 days after symptom onset20. Anti-SARS-CoV-2 mAbs which received EUA like bamlanivimab plus etesevimab and casirivimab plus imdevimab have also not been shown to be beneficial in hospitalized patients with severe COVID-19 and these products warns of worst clinical outcomes in such patients14,15. We also observed severe pneumonic changes in the group infected with a high inoculum dose treated with mAb 24 hours post infection. But the similar changes were not observed in the prophylactic and therapeutic groups exposed with a lower virus dose. Rapid induction of pneumonic changes in hamster model is a limitation in the therapeutic evaluation in the present study. The trials have also shown low clinical benefits with LY-CoV555 treatment in hospitalized COVID-19 patients25. In the present study, we have characterized ZRC 3308 mAb cocktail for COVID 19 treatment, which was found to be cross neutralizing and a promising candidate for the prophylactic use and for therapy in early cases which have not progressed to severe disease.

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   Results from TrialIdentifier: We found the following clinical trial numbers in your paper:
   

   Identifier	Status	Title
   NCT04425629	Recruiting	Safety, Tolerability, and Efficacy of Anti-Spike (S) SARS-Co…

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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 10.1101/2021.09.16.460724v1 on bioRxiv

   Sep 17, 2021