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Integrated in Silico and Experimental Approach towards the Design of a Novel Recombinant Protein Containing an Anti-HER2 scFv

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Centro de Ciências e Tecnologias Nucleares, Instituto Superior Técnico, Universidade de Lisboa, Estrada Nacional 10, ao km 139, 7, 1649-004 Lisbon, Portugal
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Faculty of Pharmacy, iMed.ULisboa—Research Institute for Medicines, University of Lisbon, 1649-004 Lisbon, Portugal
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Department of Life Sciences, University of Coimbra, Calcada Martim de Freitas, 3000-456 Coimbra, Portugal
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Center for Neuroscience and Cell Biology, Center for Inovative Biomedicine and Biotechnology, 3000-456 Coimbra, Portugal
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Departamento de Engenharia e Ciências Nucleares, Instituto Superior Técnico, Universidade de Lisboa, Estrada Nacional 10, ao km 139, 7, 1649-004 Lisbon, Portugal
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Author to whom correspondence should be addressed.
Academic Editors: Mohammad Rahimi-Gorji and Clemente Capasso
Int. J. Mol. Sci. 2021, 22(7), 3547; https://doi.org/10.3390/ijms22073547
Received: 10 February 2021 / Revised: 17 March 2021 / Accepted: 25 March 2021 / Published: 29 March 2021
(This article belongs to the Section Biochemistry)
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. View Full-Text
Keywords: cell transfection; DNA plasmid; human epidermal growth factor receptor 2; molecular docking; recombinant protein cell transfection; DNA plasmid; human epidermal growth factor receptor 2; molecular docking; recombinant protein
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MDPI and ACS Style

Santos, J.; Cardoso, M.; Moreira, I.S.; Gonçalves, J.; Correia, J.D.G.; Verde, S.C.; Melo, R. Integrated in Silico and Experimental Approach towards the Design of a Novel Recombinant Protein Containing an Anti-HER2 scFv. Int. J. Mol. Sci. 2021, 22, 3547. https://doi.org/10.3390/ijms22073547

AMA Style

Santos J, Cardoso M, Moreira IS, Gonçalves J, Correia JDG, Verde SC, Melo R. Integrated in Silico and Experimental Approach towards the Design of a Novel Recombinant Protein Containing an Anti-HER2 scFv. International Journal of Molecular Sciences. 2021; 22(7):3547. https://doi.org/10.3390/ijms22073547

Chicago/Turabian Style

Santos, Joana, Miguel Cardoso, Irina S. Moreira, João Gonçalves, João D.G. Correia, Sandra C. Verde, and Rita Melo. 2021. "Integrated in Silico and Experimental Approach towards the Design of a Novel Recombinant Protein Containing an Anti-HER2 scFv" International Journal of Molecular Sciences 22, no. 7: 3547. https://doi.org/10.3390/ijms22073547

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