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 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.
['Santos|Joana|J|', 'Cardoso|Miguel|M|', 'Moreira|Irina S|IS|0000-0003-2970-5250', 'Gonçalves|João|J|', 'Correia|João D G|JDG|0000-0002-7847-4906', 'Verde|Sandra Cabo|SC|0000-0002-6615-5289', 'Melo|Rita|R|0000-0003-1056-1007']
D003198:Computer Simulation D005822:Genetic Vectors D057809:HEK293 Cells D015700:HIV Envelope Protein gp41 D006801:Humans D062105:Molecular Docking Simulation D015202:Protein Engineering D011994:Recombinant Proteins D057127:Single-Chain Antibodies D000068878:Trastuzumab
recombinant protein DNA plasmid cell transfection human epidermal growth factor receptor 2 molecular docking