Integrated in Silico and Experimental Approach towards the Design of a Novel Recombinant Protein Containing an Anti-HER2 scFv
Biological therapies, such as recombinant proteins, are nowadays amongst the most promising approaches towards precision medicine. One of the most innovative methodologies currently available aimed at improving the production yield of recombinant proteins with minimization of costs relies on the com...
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| Format: | Article |
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MDPI AG
2021-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/7/3547 |
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| author | Joana Santos Miguel Cardoso Irina S. Moreira João Gonçalves João D. G. Correia Sandra Cabo Verde Rita Melo |
| author_facet | Joana Santos Miguel Cardoso Irina S. Moreira João Gonçalves João D. G. Correia Sandra Cabo Verde Rita Melo |
| author_sort | Joana Santos |
| collection | DOAJ |
| description | Biological therapies, such as recombinant proteins, are nowadays amongst the most promising approaches towards precision medicine. One of the most innovative methodologies currently available aimed at improving the production yield of recombinant proteins with minimization of costs relies on the combination of in silico studies to predict and deepen the understanding of the modified proteins with an experimental approach. The work described herein aims at the design and production of a biomimetic vector containing the single-chain variable domain fragment (scFv) of an anti-HER2 antibody fragment as a targeting motif fused with HIV gp41. Molecular modeling and docking studies were performed to develop the recombinant protein sequence. Subsequently, the DNA plasmid was produced and HEK-293T cells were transfected to evaluate the designed vector. The obtained results demonstrated that the plasmid construction is robust and can be expressed in the selected cell line. The multidisciplinary integrated in silico and experimental strategy adopted for the construction of a recombinant protein which can be used in HER2+-targeted therapy paves the way towards the production of other therapeutic proteins in a more cost-effective way. |
| first_indexed | 2024-03-10T12:48:17Z |
| format | Article |
| id | doaj.art-4ce5efdc3adf4839b331f00e6cbf8f2f |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T12:48:17Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-4ce5efdc3adf4839b331f00e6cbf8f2f2023-11-21T13:19:21ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-03-01227354710.3390/ijms22073547Integrated in Silico and Experimental Approach towards the Design of a Novel Recombinant Protein Containing an Anti-HER2 scFvJoana Santos0Miguel Cardoso1Irina S. Moreira2João Gonçalves3João D. G. Correia4Sandra Cabo Verde5Rita Melo6Centro de Ciências e Tecnologias Nucleares, Instituto Superior Técnico, Universidade de Lisboa, Estrada Nacional 10, ao km 139, 7, 1649-004 Lisbon, PortugalFaculty of Pharmacy, iMed.ULisboa—Research Institute for Medicines, University of Lisbon, 1649-004 Lisbon, PortugalDepartment of Life Sciences, University of Coimbra, Calcada Martim de Freitas, 3000-456 Coimbra, PortugalFaculty of Pharmacy, iMed.ULisboa—Research Institute for Medicines, University of Lisbon, 1649-004 Lisbon, PortugalCentro de Ciências e Tecnologias Nucleares, Instituto Superior Técnico, Universidade de Lisboa, Estrada Nacional 10, ao km 139, 7, 1649-004 Lisbon, PortugalCentro de Ciências e Tecnologias Nucleares, Instituto Superior Técnico, Universidade de Lisboa, Estrada Nacional 10, ao km 139, 7, 1649-004 Lisbon, PortugalCentro de Ciências e Tecnologias Nucleares, Instituto Superior Técnico, Universidade de Lisboa, Estrada Nacional 10, ao km 139, 7, 1649-004 Lisbon, PortugalBiological therapies, such as recombinant proteins, are nowadays amongst the most promising approaches towards precision medicine. One of the most innovative methodologies currently available aimed at improving the production yield of recombinant proteins with minimization of costs relies on the combination of in silico studies to predict and deepen the understanding of the modified proteins with an experimental approach. The work described herein aims at the design and production of a biomimetic vector containing the single-chain variable domain fragment (scFv) of an anti-HER2 antibody fragment as a targeting motif fused with HIV gp41. Molecular modeling and docking studies were performed to develop the recombinant protein sequence. Subsequently, the DNA plasmid was produced and HEK-293T cells were transfected to evaluate the designed vector. The obtained results demonstrated that the plasmid construction is robust and can be expressed in the selected cell line. The multidisciplinary integrated in silico and experimental strategy adopted for the construction of a recombinant protein which can be used in HER2+-targeted therapy paves the way towards the production of other therapeutic proteins in a more cost-effective way.https://www.mdpi.com/1422-0067/22/7/3547cell transfectionDNA plasmidhuman epidermal growth factor receptor 2molecular dockingrecombinant protein |
| spellingShingle | Joana Santos Miguel Cardoso Irina S. Moreira João Gonçalves João D. G. Correia Sandra Cabo Verde Rita Melo Integrated in Silico and Experimental Approach towards the Design of a Novel Recombinant Protein Containing an Anti-HER2 scFv International Journal of Molecular Sciences cell transfection DNA plasmid human epidermal growth factor receptor 2 molecular docking recombinant protein |
| title | Integrated in Silico and Experimental Approach towards the Design of a Novel Recombinant Protein Containing an Anti-HER2 scFv |
| title_full | Integrated in Silico and Experimental Approach towards the Design of a Novel Recombinant Protein Containing an Anti-HER2 scFv |
| title_fullStr | Integrated in Silico and Experimental Approach towards the Design of a Novel Recombinant Protein Containing an Anti-HER2 scFv |
| title_full_unstemmed | Integrated in Silico and Experimental Approach towards the Design of a Novel Recombinant Protein Containing an Anti-HER2 scFv |
| title_short | Integrated in Silico and Experimental Approach towards the Design of a Novel Recombinant Protein Containing an Anti-HER2 scFv |
| title_sort | integrated in silico and experimental approach towards the design of a novel recombinant protein containing an anti her2 scfv |
| topic | cell transfection DNA plasmid human epidermal growth factor receptor 2 molecular docking recombinant protein |
| url | https://www.mdpi.com/1422-0067/22/7/3547 |
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