Engineered Human Monoclonal scFv to Receptor Binding Domain of Ebolavirus
Abstract
1. Introduction
2. Materials and Methods
2.1. Culture Media, Cells, Antibodies, and Reagents Used in This Study
2.2. Plasmid Constructs
2.3. Production of Recombinant EBOV Receptor-Binding Domain (RBD)
2.4. Production of Human Single-Chain Antibody Variable Fragments (HuscFvs) That Bind to the EBOV RBD
2.5. Large Scale Production of the HuscFvs and Generation of Cell-Penetrating HuscFvs
2.6. Western Blotting
2.7. Biocompatibility of R9-HuscFvs/HuscFvs to Mammalian Cells
2.8. Determination of Cell-Penetrating Ability of R9-HuscFvs
2.9. Preparation of EBOV-Like Particles and Transmission Electron Microscopy
2.10. Determination of the Ability of the R9-HuscFvs/HuscFvs to EBOV RBD in Inhibiting Cellular Entry of the VLP
2.11. Preparation of Cells That Exposed EBOV GP-RBD
2.12. Determination of Binding of R9-HuscFvs/HuscFvs to RBD-Exposed Cells
2.13. Protein Modeling and Molecular Docking
2.14. Statistical Analysis
3. Results
3.1. Recombinant EBOV GP-RBD and HuscFvs to the RBD
3.2. Cell-Penetrating HuscFvs and Their Biocompatibility to Human Cells
3.3. Ability of the R9-HuscFvs/HuscFvs in Inhibiting Cellular Entry of the Ebola VLP
3.4. Binding of the R9-HuscFvs/HuscFvs to RBD
3.5. Computerized Simulation of HuscFv-RBD Interaction
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Densumite, J.; Phanthong, S.; Seesuay, W.; Sookrung, N.; Chaisri, U.; Chaicumpa, W. Engineered Human Monoclonal scFv to Receptor Binding Domain of Ebolavirus. Vaccines 2021, 9, 457. https://doi.org/10.3390/vaccines9050457
Densumite J, Phanthong S, Seesuay W, Sookrung N, Chaisri U, Chaicumpa W. Engineered Human Monoclonal scFv to Receptor Binding Domain of Ebolavirus. Vaccines. 2021; 9(5):457. https://doi.org/10.3390/vaccines9050457
Chicago/Turabian StyleDensumite, Jaslan, Siratcha Phanthong, Watee Seesuay, Nitat Sookrung, Urai Chaisri, and Wanpen Chaicumpa. 2021. "Engineered Human Monoclonal scFv to Receptor Binding Domain of Ebolavirus" Vaccines 9, no. 5: 457. https://doi.org/10.3390/vaccines9050457
APA StyleDensumite, J., Phanthong, S., Seesuay, W., Sookrung, N., Chaisri, U., & Chaicumpa, W. (2021). Engineered Human Monoclonal scFv to Receptor Binding Domain of Ebolavirus. Vaccines, 9(5), 457. https://doi.org/10.3390/vaccines9050457