Potent Neutralization of SARS-CoV-2 by Hetero-Bivalent Alpaca Nanobodies Targeting the Spike Receptor-Binding Domain
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Abstract
To date, SARS-CoV-2 has caused tremendous loss of human life and economic output worldwide. Although a few COVID-19 vaccines have been approved in several countries, the development of effective therapeutics, including SARS-CoV-2 targeting antibodies, remains critical.
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SciScore for 10.1101/2021.02.02.429311: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: Phage display library construction: The experiments involved alpacas were approved by a local ethics committee. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Two female alpacas were immunized by 2 times of subcutaneous injection and 1 time of intramuscular injection, each with 500 μg SARS-CoV-2 RBD in PBS, which was emulsified with an equal volume of Freund’s adjuvant (Sigma Aldrich). Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources After incubation in RBD coated wells and standard washing, bound Nb-Fc was detected with a monoclonal anti-IgG1 Fc-HRP antibody. anti-IgG1 …SciScore for 10.1101/2021.02.02.429311: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: Phage display library construction: The experiments involved alpacas were approved by a local ethics committee. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Two female alpacas were immunized by 2 times of subcutaneous injection and 1 time of intramuscular injection, each with 500 μg SARS-CoV-2 RBD in PBS, which was emulsified with an equal volume of Freund’s adjuvant (Sigma Aldrich). Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources After incubation in RBD coated wells and standard washing, bound Nb-Fc was detected with a monoclonal anti-IgG1 Fc-HRP antibody. anti-IgG1 Fc-HRPsuggested: (SouthernBiotech Cat# 9054-05, RRID:AB_2796627)After 2 days of infection, the cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton-100, blocked with DMEM containing 10% FBS, and stained with a rabbit monoclonal antibody against SARS-CoV-2 NP (GeneTex, GTX635679) and an Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (ThermoFisher Scientific). SARS-CoV-2 NPsuggested: NoneGTX635679suggested: (GeneTex Cat# GTX635679, RRID:AB_2888553)anti-mousesuggested: NoneExperimental Models: Cell Lines Sentences Resources The expression vectors were transiently transfected to human HEK293F cells with polyethylenimine (Polyscience). HEK293Fsuggested: RRID:CVCL_6642)The antibody and virus mixture were then added to Vero E6 cells in 96-well plates (Corning). Vero E6suggested: NoneSoftware and Algorithms Sentences Resources Each experiment was repeated twice, and the data were fitted with Prism to obtain the Tm values. Prismsuggested: (PRISM, RRID:SCR_005375)The total number of cells, as indicated by the nuclei staining, and the infected cells, as indicated by the NP staining, were quantified with the cellular analysis module of the Gen5 software (BioTek). Gen5suggested: (Gen5, RRID:SCR_017317)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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