Integrating Biochemical and Computational Approaches Reveal Structural Insights in Trastuzumab scFv-Fc Antibody Engineering
Antibody-based agents have become a preferred treatment for various diseases, including cancer, due to significant advances in antibody engineering. The use of single-chain Fv-Fcs (scFv-Fcs) has been a promising engineering approach for therapeutic design. The concept is that the Fc provides increas...
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MDPI AG
2025-04-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/15/5/606 |
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| author | Olga Bednova Jessica Pougoue Ketchemen Hazem Mslati Mark Barok Heikki Joensuu Natalie Zeytuni Francesco Gentile Leon Sanche Humphrey Fonge Jeffrey Victor Leyton |
| author_facet | Olga Bednova Jessica Pougoue Ketchemen Hazem Mslati Mark Barok Heikki Joensuu Natalie Zeytuni Francesco Gentile Leon Sanche Humphrey Fonge Jeffrey Victor Leyton |
| author_sort | Olga Bednova |
| collection | DOAJ |
| description | Antibody-based agents have become a preferred treatment for various diseases, including cancer, due to significant advances in antibody engineering. The use of single-chain Fv-Fcs (scFv-Fcs) has been a promising engineering approach for therapeutic design. The concept is that the Fc provides increased stability and target binding and ultimately improves performance. However, the structural and dynamic relationship between the variable and Fc domains, which are fused in close proximity, and the impact on stability and target binding are not well understood. This study evaluated trastuzumab-derived scFv-Fc antibodies, focusing on the impact of their design on important biopharmaceutical parameters. Computational modelling and molecular dynamics, alongside experimental studies, were used to ascertain their dynamics, expression and purification, stabilities, and binding potencies. The results showed that the scFv subunits exhibited stochastic interplays that lead to diverse shapes and were associated with functional performance. This new understanding of scFv-Fc antibodies and their structural and functional nuances provides important details to further guide the design of more effective and less toxic therapeutics. |
| format | Article |
| id | doaj-art-b5a61a33db6343fc82a18d6240c84585 |
| institution | OA Journals |
| issn | 2218-273X |
| language | English |
| publishDate | 2025-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj-art-b5a61a33db6343fc82a18d6240c845852025-08-20T01:56:29ZengMDPI AGBiomolecules2218-273X2025-04-0115560610.3390/biom15050606Integrating Biochemical and Computational Approaches Reveal Structural Insights in Trastuzumab scFv-Fc Antibody EngineeringOlga Bednova0Jessica Pougoue Ketchemen1Hazem Mslati2Mark Barok3Heikki Joensuu4Natalie Zeytuni5Francesco Gentile6Leon Sanche7Humphrey Fonge8Jeffrey Victor Leyton9Département de Médecine Nucléaire et Radiobiologie, Faculté de Médecine et des Sciences de la Santé, Université de Sherbrooke, Sherbrooke, QC J1H 5N4, CanadaFaculté de Pharmacie, Université Laval, Québec, QC G1V 0A6, CanadaDepartment of Cellular and Molecular Medicine, Faculty of Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada Helsinki University Hospital and University of Helsinki, 00290 Helsinki, Finland Helsinki University Hospital and University of Helsinki, 00290 Helsinki, FinlandDepartment of Anatomy and Cell Biology, McGill University, Montreal, QC H3A 0C7, CanadaDepartment of Chemistry and Biomedical Sciences, University of Ottawa, Ottawa, ON K1H 8M5, CanadaDépartement de Médecine Nucléaire et Radiobiologie, Faculté de Médecine et des Sciences de la Santé, Université de Sherbrooke, Sherbrooke, QC J1H 5N4, CanadaFaculté de Pharmacie, Université Laval, Québec, QC G1V 0A6, CanadaDepartment of Cellular and Molecular Medicine, Faculty of Medicine, University of Ottawa, Ottawa, ON K1H 8M5, CanadaAntibody-based agents have become a preferred treatment for various diseases, including cancer, due to significant advances in antibody engineering. The use of single-chain Fv-Fcs (scFv-Fcs) has been a promising engineering approach for therapeutic design. The concept is that the Fc provides increased stability and target binding and ultimately improves performance. However, the structural and dynamic relationship between the variable and Fc domains, which are fused in close proximity, and the impact on stability and target binding are not well understood. This study evaluated trastuzumab-derived scFv-Fc antibodies, focusing on the impact of their design on important biopharmaceutical parameters. Computational modelling and molecular dynamics, alongside experimental studies, were used to ascertain their dynamics, expression and purification, stabilities, and binding potencies. The results showed that the scFv subunits exhibited stochastic interplays that lead to diverse shapes and were associated with functional performance. This new understanding of scFv-Fc antibodies and their structural and functional nuances provides important details to further guide the design of more effective and less toxic therapeutics.https://www.mdpi.com/2218-273X/15/5/606trastuzumabantibody engineeringHER2scFv-Fcmolecular dynamic simulation |
| spellingShingle | Olga Bednova Jessica Pougoue Ketchemen Hazem Mslati Mark Barok Heikki Joensuu Natalie Zeytuni Francesco Gentile Leon Sanche Humphrey Fonge Jeffrey Victor Leyton Integrating Biochemical and Computational Approaches Reveal Structural Insights in Trastuzumab scFv-Fc Antibody Engineering Biomolecules trastuzumab antibody engineering HER2 scFv-Fc molecular dynamic simulation |
| title | Integrating Biochemical and Computational Approaches Reveal Structural Insights in Trastuzumab scFv-Fc Antibody Engineering |
| title_full | Integrating Biochemical and Computational Approaches Reveal Structural Insights in Trastuzumab scFv-Fc Antibody Engineering |
| title_fullStr | Integrating Biochemical and Computational Approaches Reveal Structural Insights in Trastuzumab scFv-Fc Antibody Engineering |
| title_full_unstemmed | Integrating Biochemical and Computational Approaches Reveal Structural Insights in Trastuzumab scFv-Fc Antibody Engineering |
| title_short | Integrating Biochemical and Computational Approaches Reveal Structural Insights in Trastuzumab scFv-Fc Antibody Engineering |
| title_sort | integrating biochemical and computational approaches reveal structural insights in trastuzumab scfv fc antibody engineering |
| topic | trastuzumab antibody engineering HER2 scFv-Fc molecular dynamic simulation |
| url | https://www.mdpi.com/2218-273X/15/5/606 |
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