A chicken IgY can efficiently inhibit the entry and replication of SARS-CoV-2 by targeting the ACE2 binding domain in vitro
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Abstract
COVID-19 pneumonia has now spread widely in the world. Currently, no specific antiviral drugs are available. The vaccine is the most effective way to control the epidemic. Passive immune antibodies are also an effective method to prevent and cure COVID-19 pneumonia. We used the SARS-CoV-2 S receptor-binding domain (RBD) as an antigen to immunize layers in order to extract, separate, and purify SARS-CoV-2-IgY from egg yolk. SARS-CoV-2-IgY (S-IgY)can block the entry of SARS-CoV-2 into the Cells and reduce the viral load in cells. The Half effective concentration (EC 50 ) of W3-IgY (S-IgY in the third week after immunization) is 1.35 ± 0.15nM. The EC 50 of W9-IgY (S-IgY in the ninth week after immunization) is 2.76 ± 1.54 nM. When the dose of S-IgY is 55 nM, the fluorescence representing intracellular viral protein is obviously weakened in Immunofluorescence microscopy.
Results of Sars-CoV-2 /Vero E6 cell experiment confirmed that S-IgY has a strong antiviral effect on SARS-CoV-2, and its (EC 50 ) is 27.78 ±1.54 nM vs 3,259 ± 159.62 nM of Redesivir (differ > 106 times P<0 . 001 ).
S-IgY can inhibit the entry and replication of SARS-CoV-2, which is related to its targeting the ACE2 binding domain.
S-IgY is safe, efficient, stable, and easy to obtain. This antibody can be an effective tool for preventing and treating COVID-19 pneumonia.
Abstract Figure
Fig. 1.Graphical Abstract
The figure briefly illustrates that the preparation and extraction of S-IgY and its anti-S-CoV-2 mechanism is to inhibit the entry and replication of SARS-CoV-2 by targeting the ACE2.
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SciScore for 10.1101/2021.02.16.430255: (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 second antibody was prepared by diluting HRP-labeled goat anti-chicken(Provided by ZSGB-Bio) with a ratio of 1:10000, and incubated for 1.5 h at room temperature, TBST washed the membrane for 5 times, The mixed immunoblotting chemiluminescence solution was dripped on NC membrane, incubated at room temperature for 2-3 minutes, then placed in a cassette, and the X-ray film was developed (See Fig.4). 6. anti-chicken(Provided by ZSGB-Biosuggested: NoneThe … SciScore for 10.1101/2021.02.16.430255: (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 second antibody was prepared by diluting HRP-labeled goat anti-chicken(Provided by ZSGB-Bio) with a ratio of 1:10000, and incubated for 1.5 h at room temperature, TBST washed the membrane for 5 times, The mixed immunoblotting chemiluminescence solution was dripped on NC membrane, incubated at room temperature for 2-3 minutes, then placed in a cassette, and the X-ray film was developed (See Fig.4). 6. anti-chicken(Provided by ZSGB-Biosuggested: NoneThe cells were further incubated with the primary antibody (a monoclonal antibody against viral S protein) for 2 h, followed by incubation with the secondary antibody (Alexa 488-labeled goat anti-mouse[1:500; Abcam]). anti-mouse[1:500suggested: NoneExperimental Models: Cell Lines Sentences Resources Forty-eight hours posttransfection, 150 μl pseudotyped VSV-ΔG bearing VSV-G protein were used to infect Vero E6 cells. Vero E6suggested: RRID:CVCL_XD71)Experimental Models: Organisms/Strains Sentences Resources SARS-CoV-2 -IgY (dilution ratio: 1:10000) was incubated overnight by inversion method. SARS-CoV-2 -IgYsuggested: NoneSoftware and Algorithms Sentences Resources Western blot: SARS-CoV-2 S-RBD (provided by Sina Biological Company) was separated by polyacrylamide amine gel (10% separation gel+5% concentration gel) under the electrophoresis condition of 50 mA for 60 min. Sinasuggested: (SINA, RRID:SCR_005067)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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