The oxidized low-density lipoprotein receptor-1 (LOX-1) targeted single-chain variable fragment (scFvs) is a promising molecule\nfor the targeted delivery of imaging and therapeutic molecules of atherosclerotic diseases; however, its applications are limited by\nthe inherent low antigen affinity. In this study, the three-dimensional (3D) model of the anti-LOX-1 scFv was constructed and its\ndocking with the LOX-1 protein was developed. To improve the LOX-1-binding activity, the anti-LOX-1 scFv was designed to fuse\nwith one of three LOX-1-binding heptapeptides, LTPATAI, FQTPPQL, and LSIPPKA, at its N-terminus and C-terminus and in\nthe linker region, which have different LOX-1-binding interfaces with the anti-LOX-1 scFv analyzed by an array of computational\napproaches. These scFv/peptide fusions were constructed, successfully expressed in Brevibacillus choshinensis hosts, and purified\nby a two-step column purification process. The antigen binding activity, structural characteristics, thermal stability, and stability in\nserum of these fusion proteins were examined. Results showed that the scFv with N-terminal fusing peptides proteins demonstrated\nincreased LOX-1-binding activity without decrease in stability. These findings will help increase the application efficacy of LOX-1\ntargeting scFv in LOX-1-based therapy.
Loading....