Author(s):
Santos, Joana ; Cardoso, Miguel ; Moreira, Irina S. ; Gonçalves, João ; Correia, João D. G. ; Verde, Sandra Cabo ; Melo, Rita
Date: 2021
Persistent ID: https://hdl.handle.net/10316/104754
Origin: Estudo Geral - Universidade de Coimbra
Subject(s): cell transfection; DNA plasmid; human epidermal growth factor receptor 2; molecular docking; recombinant protein; Computer Simulation; Genetic Vectors; HEK293 Cells; HIV Envelope Protein gp41; Humans; Molecular Docking Simulation; Protein Engineering; Recombinant Proteins; Single-Chain Antibodies; Trastuzumab
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.
