Synthetic antibodies neutralize SARS-CoV-2 infection of mammalian cells

Abstract

Coronaviruses (CoV) are a large family of enveloped, RNA viruses that circulate in mammals and birds. Three highly pathogenic strains have caused zoonotic infections in humans that result in severe respiratory syndromes including the Middle East Respiratory Syndrome CoV (MERS), Severe Acute Respiratory Syndrome CoV (SARS), and the ongoing Coronavirus Disease 2019 (COVID-19) pandemic. Here, we describe a panel of synthetic monoclonal antibodies, built on a human IgG framework, that bind to the spike protein of SARS-CoV-2 (the causative agent of COVID-19), compete for ACE2 binding, and potently inhibit SARS-CoV-2. All antibodies that exhibited neutralization potencies at sub-nanomolar concentrations against SARS-CoV-2/USA/WA1 in Vero E6 cells, also bound to the receptor binding domain (RBD), suggesting competition for the host receptor ACE2. These antibodies represent strong immunotherapeutic candidates for treatment of COVID-19.

SciScore for 10.1101/2020.06.05.137349: (What is this?)

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

Table 1: Rigor

Institutional Review Board Statement	not detected.
Randomization	not detected.
Blinding	not detected.
Power Analysis	not detected.
Sex as a biological variable	not detected.
Cell Line Authentication	not detected.

Table 2: Resources

Antibodies
Sentences	Resources
The plates were washed with PBST and incubated for 30 min with anti-k-HRP antibody conjugate (1:7500 dilution in PBT).	anti-k-HRP suggested: None
Plates were washed and sequentially incubated with 1 µg/mL anti-S antibody CR3022 (17) and HRP-conjugated goat anti-human IgG in PBS supplemented with 0.1% saponin and 0.1% BSA.	anti-S suggested: None anti-human IgG suggested: None
Experimental Models: Cell Lines
Sentences	Resources
The ACE2-expressing cell line was …

SciScore for 10.1101/2020.06.05.137349: (What is this?)

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

Table 1: Rigor

Institutional Review Board Statement	not detected.
Randomization	not detected.
Blinding	not detected.
Power Analysis	not detected.
Sex as a biological variable	not detected.
Cell Line Authentication	not detected.

Table 2: Resources

Antibodies
Sentences	Resources
The plates were washed with PBST and incubated for 30 min with anti-k-HRP antibody conjugate (1:7500 dilution in PBT).	anti-k-HRP suggested: None
Plates were washed and sequentially incubated with 1 µg/mL anti-S antibody CR3022 (17) and HRP-conjugated goat anti-human IgG in PBS supplemented with 0.1% saponin and 0.1% BSA.	anti-S suggested: None anti-human IgG suggested: None
Experimental Models: Cell Lines
Sentences	Resources
The ACE2-expressing cell line was constructed by co-transfecting the Freestyle 293-F cells with the PB-CMV plasmid encoding the full length human ACE2 and the pCyL43 plasmid.	293-F suggested: None
Vectors for the heavy and light chains were transfected into HEK293F cells (Invitrogen, Grand Island, NY) using FectoPro according to the manufacturer’s instructions (Polyplus Transfection, NY).	HEK293F suggested: RRID:CVCL_6642)
Virus stocks were produced in Vero CCL81 cells (ATCC) and titrated by focus-forming assay on Vero E6 cells.	Vero CCL81 suggested: None Vero E6 suggested: RRID:CVCL_XD71)
Software and Algorithms
Sentences	Resources
To estimate EC50 values, data were fit to standard four-parameter logistic equations using Graphpad Prism (GraphPad Software, La Jolla, CA).	Graphpad Prism suggested: (GraphPad Prism, RRID:SCR_002798)
Data were processed using Prism software (GraphPad Prism 8.0).	Prism suggested: (PRISM, RRID:SCR_005375) GraphPad suggested: (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.

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Synthetic antibodies neutralize SARS-CoV-2 infection of mammalian cells

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