Development of Equine Immunoglobulin Fragment F(ab’) 2 with High Neutralizing Capability against SARS-CoV-2
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Abstract
The ongoing pandemic, COVID-19, caused by SARS-CoV-2 has taken the world, and especially the scientific community by storm. While vaccines are being introduced into the market, there is also a pressing need to find potential drugs and therapeutic modules. Remdesivir is one of the antivirals currently being used with a limited window of action. As more drugs are being vetted, passive immunotherapy in the form of neutralizing antibodies can provide immediate action to combat the increasing numbers of COVID-positive cases. Herein, we demonstrate that equines hyper-immunized with chemically inactivated SARS-CoV-2 generate high titers of antibody with a strong virus neutralizing potential. ELISA performed with pooled antisera displayed highest immunoglobulin titer on 42 days post-immunization, at 1:51,200 dilutions. F(ab’) 2 immunoglobulin fragments generated from the pools also showed very high, antigen-specific affinity at 1:102,400 dilutions. Finally, in vitro virus neutralization assays confirmed that different pools of F(ab’) 2 fragments could successfully neutralize SARS-CoV-2 with titers well above 25,000, indicating the potential of this strategy in treating severe COVID-19 cases with high titers. The F(ab’) 2 was able to cross neutralize another SARS-CoV-2 strain, demonstrating its efficacy against the emerging viral variants and the importance of this approach in our efforts of eradication of COVID-19. In conclusion, this study demonstrates that virus-neutralizing antibodies raised in equines can potentially be used as a treatment regimen in the form of effective passive immunotherapy to combat COVID-19.
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SciScore for 10.1101/2021.03.09.434030: (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 Plasma samples from the immunized animals were tested periodically to estimate the antibody response against SARS-CoV-2 inactivated viral antigen. SARS-CoV-2 inactivated viral antigen.suggested: NoneSubsequently, the plate was washed four times and incubated with HRP conjugated anti-horse whole IgG secondary antibody (Sigma) for 1 hr at RT. anti-horse whole IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources 100μL of the filter-sterilized VTM … SciScore for 10.1101/2021.03.09.434030: (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 Plasma samples from the immunized animals were tested periodically to estimate the antibody response against SARS-CoV-2 inactivated viral antigen. SARS-CoV-2 inactivated viral antigen.suggested: NoneSubsequently, the plate was washed four times and incubated with HRP conjugated anti-horse whole IgG secondary antibody (Sigma) for 1 hr at RT. anti-horse whole IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources 100μL of the filter-sterilized VTM was added to Vero cell monolayer in 96 well plates. Verosuggested: NoneSoftware and Algorithms Sentences Resources Image processing was performed using ImageJ22. ImageJ22suggested: NoneResults 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 found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
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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