Major role of IgM in the neutralizing activity of convalescent plasma against SARS-CoV-2
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SciScore for 10.1101/2020.10.09.333278: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: Ethics statement: All work was conducted in accordance with the Declaration of Helsinki in terms of informed consent and approval by an appropriate Ethics Review board.
IRB: Ethics statement: All work was conducted in accordance with the Declaration of Helsinki in terms of informed consent and approval by an appropriate Ethics Review board.Randomization not detected. Blinding The assay was conducted with the person blinded to the sample identity. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Isotype depletion: Selective depletion of IgM, IgA or IgG was … SciScore for 10.1101/2020.10.09.333278: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: Ethics statement: All work was conducted in accordance with the Declaration of Helsinki in terms of informed consent and approval by an appropriate Ethics Review board.
IRB: Ethics statement: All work was conducted in accordance with the Declaration of Helsinki in terms of informed consent and approval by an appropriate Ethics Review board.Randomization not detected. Blinding The assay was conducted with the person blinded to the sample identity. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Isotype depletion: Selective depletion of IgM, IgA or IgG was done by adsorption on isotype-specific ligands immobilized on sepharose or agarose beads starting with a five-fold dilution of plasma in PBS. IgG and IgA antibodies were depleted from plasma obtained from 25 recovered COVID-19 patient using Protein G HP Spintrap (GE Healthcare Life Sciences, Buckinghamshire, UK) and Peptide M / Agarose (InvivoGen, San Diego, CA), respectively, according to the manufacturer’s instructions with the exception that no elution step for the recovery of the targeted antibodies was done. IgAsuggested: NonePlasma binding to cell-surface Spike was revealed using fluorescent secondary antibodies able to detect all Ig isotypes (anti-human IgM+IgG+IgA; Jackson ImmunoResearch Laboratories, Inc.) or specific to IgG isotype (Biolegend), IgM isotype (Jackson ImmunoResearch Laboratories, Inc.) or IgA isotype (Jackson ImmunoResearch Laboratories, Inc.). anti-human IgM+IgG+IgAsuggested: NoneIgG isotype (Biolegend), IgMsuggested: NoneIgM isotypesuggested: NoneIgA isotype (Jackson ImmunoResearch Laboratories, Inc.)suggested: NoneAn anti-mouse SARS-CoV-2 nucleocapsid protein (Clone 1C7, Bioss Antibodies) primary antibody solution was prepared at 1 μg/mL in PBS + 1% non-fat milk and added to all wells for one hour at room temperature. anti-mouse SARS-CoV-2 nucleocapsid proteinsuggested: NoneFollowing extensive washing with PBS, an anti-mouse IgG HRP secondary antibody solution was formulated in PBS + 1% non-fat milk. anti-mouse IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources The 293T-ACE2 cell line was previously reported (Prévost et al., 2020). 293T-ACE2suggested: RRID:CVCL_YZ65)Supernatant containing lentiviral particles was used to infect 293T cells in presence of 5μg/mL polybrene. 293Tsuggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)One day prior to infection, 2×104 Vero E6 cells were seeded per well of a 96 well flat bottom plate and incubated overnight (37°C/5% CO2) to permit Vero E6 cell adherence. Vero E6suggested: NoneSoftware and Algorithms Sentences Resources After four washes, 100 μL of either goat anti-human IgA+G+M (H+L) HRP conjugate (1/30 000) for the detection of all isotypes, goat anti-human IgM (μ-chain specific) HRP conjugate (1/15 000), F(ab’)₂ fragment goat anti-human IgA (α-chain specific) HRP conjugate (1/4500) (all from Jackson Immunosearch Laboratories, Inc.) or goat anti-human IgG (γ-chain specific) HRP conjugate (1/50 000) (Invitrogen) were added and samples were incubated at 20-24°C for 60 minutes. Jackson Immunosearch Laboratoriessuggested: NoneSamples were acquired on a LSRII cytometer (BD Biosciences, Mississauga, ON, Canada) and data analysis was performed using FlowJo v10.5.3 (Tree Star, Ashland, OR). FlowJosuggested: (FlowJo, RRID:SCR_008520)Statistical analysis: Statistics were analyzed using GraphPad Prism version 8.0.2 (GraphPad, San Diego, CA, (USA). GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)GraphPadsuggested: (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.
- 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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