Centro de Ciências e Tecnologias Nucleares, Instituto Superior Técnico, Universidade de Lisboa, Estrada Nacional 10, ao km 139, 7, 1649-004 Lisbon, Portugal.
Faculty of Pharmacy, iMed.ULisboa-Research Institute for Medicines, University of Lisbon, 1649-004 Lisbon, Portugal.
Int J Mol Sci. 2021 Mar 29;22(7):3547. doi: 10.3390/ijms22073547.
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.
生物疗法，如重组蛋白，是当今最有前途的精准医学方法之一。目前，最具创新性的方法之一是结合计算机模拟研究，预测和深入了解经过修饰的蛋白质，从而提高重组蛋白的产量并降低成本。本文旨在设计和生产一种仿生载体，该载体包含抗 HER2 抗体片段的单链可变区片段 (scFv) 作为靶向结构域，与 HIV gp41 融合。进行了分子建模和对接研究以开发重组蛋白序列。随后，生产了 DNA 质粒并转染 HEK-293T 细胞以评估设计的载体。所得结果表明，质粒构建是稳健的，可以在选定的细胞系中表达。用于构建可用于 HER2+靶向治疗的重组蛋白的多学科集成计算和实验策略为以更具成本效益的方式生产其他治疗性蛋白铺平了道路。
