A novel biparatopic antibody-ACE2 fusion that blocks SARS-CoV-2 infection: implications for therapy
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Abstract
In the absence of a proven effective vaccine preventing infection by SARS-CoV-2, or a proven drug to treat COVID-19, the positive results of passive immune therapy using convalescent serum provides a strong lead. We have developed a new class of tetravalent, biparatopic therapy, 89C8-ACE2. It combines the specificity of a monoclonal antibody (89C8) that recognizes the relatively conserved N-terminal domain (NTD) of the viral S glycoprotein, and the ectodomain of ACE2, which binds to the receptor-binding domain (RBD) of S. This molecule shows exceptional performance in vitro, inhibiting the interaction of recombinant S1 to ACE2 and transduction of ACE2-overexpressing cells by S-pseudotyped lentivirus with IC50s substantially below 100 pM, and with potency approximately 100-fold greater than ACE2-Fc itself. Moreover, 89C8-ACE2 was able to neutralize authentic virus infection in a standard assay at low nanomolar concentrations, making this class of molecule a promising lead for therapeutic applications.
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SciScore for 10.1101/2020.06.14.147868: (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 Recombinant antibody expression: The cDNAs encoding the variable regions of heavy (human IgG1 heavy chain) and light (kappa light chain constant regions) chains were cloned into expression plasmids (pCDNA3.1). human IgG1 heavy chainsuggested: NoneCell based blocking test: Gradient diluted antibodies or antibody-ACE2 fusion proteins were first pre-incubated with 0.1nM S1-mFc (SinoBiological) at 37 °C overnight, followed by incubation with CHO cells stably … SciScore for 10.1101/2020.06.14.147868: (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 Recombinant antibody expression: The cDNAs encoding the variable regions of heavy (human IgG1 heavy chain) and light (kappa light chain constant regions) chains were cloned into expression plasmids (pCDNA3.1). human IgG1 heavy chainsuggested: NoneCell based blocking test: Gradient diluted antibodies or antibody-ACE2 fusion proteins were first pre-incubated with 0.1nM S1-mFc (SinoBiological) at 37 °C overnight, followed by incubation with CHO cells stably expressing ACE2 protein (CHO-ACE2) for 1h at RT. antibody-ACE2suggested: NoneExperimental Models: Cell Lines Sentences Resources The heavy- and light-chain encoding plasmids were transiently co-transfected into Expi293 cells (Life technologies) using PEI. Expi293suggested: RRID:CVCL_D615)Cell based blocking test: Gradient diluted antibodies or antibody-ACE2 fusion proteins were first pre-incubated with 0.1nM S1-mFc (SinoBiological) at 37 °C overnight, followed by incubation with CHO cells stably expressing ACE2 protein (CHO-ACE2) for 1h at RT. CHOsuggested: CLS Cat# 603479/p746_CHO, RRID:CVCL_0213)HEK293T overexpressing hACE2 were seeded into the 96-well plates 16h before infection. HEK293Tsuggested: NoneThereafter, 0.5 mL of a single cell suspension of Vero E6 cells in D1 at 5 E5/mL was added, and incubated for 2 h at 37 °C before being overlain with 0.5 mL of D1 supplemented with carboxymethyl cellulose (1.5 %). Vero E6suggested: NoneSoftware and Algorithms Sentences Resources The IC50 values were calculated with non-linear regression log (inhibitor) vs. response (four parameters) using GraphPad Prism 7 (GraphPad Software, Inc.) GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)The PRNT90 and PRNT50 values were calculated using the “FindECanything” non-linear curve fitting procedure of GraphPad Prism 8.4.1, with F set to 10 and 50, respectively. 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.
- 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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