Improved scFv Anti-LOX-1 Binding Activity by Fusion with LOX-1-Binding Peptides
The oxidized low-density lipoprotein receptor-1 (LOX-1) targeted single-chain variable fragment (scFvs) is a promising molecule for the targeted delivery of imaging and therapeutic molecules of atherosclerotic diseases; however, its applications are limited by the inherent low antigen affinity. In t...
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doaj-f4fc2de30626407884322d40488b361d2020-11-25T02:46:15ZengHindawi LimitedBioMed Research International2314-61332314-61412017-01-01201710.1155/2017/89469358946935Improved scFv Anti-LOX-1 Binding Activity by Fusion with LOX-1-Binding PeptidesWei Hu0Qiuhong Xie1Hongyu Xiang2School of Life Science, Jilin University, Changchun, Jilin 130012, ChinaSchool of Life Science, Jilin University, Changchun, Jilin 130012, ChinaSchool of Life Science, Jilin University, Changchun, Jilin 130012, ChinaThe oxidized low-density lipoprotein receptor-1 (LOX-1) targeted single-chain variable fragment (scFvs) is a promising molecule for the targeted delivery of imaging and therapeutic molecules of atherosclerotic diseases; however, its applications are limited by the inherent low antigen affinity. In this study, the three-dimensional (3D) model of the anti-LOX-1 scFv was constructed and its docking with the LOX-1 protein was developed. To improve the LOX-1-binding activity, the anti-LOX-1 scFv was designed to fuse with one of three LOX-1-binding heptapeptides, LTPATAI, FQTPPQL, and LSIPPKA, at its N-terminus and C-terminus and in the linker region, which have different LOX-1-binding interfaces with the anti-LOX-1 scFv analyzed by an array of computational approaches. These scFv/peptide fusions were constructed, successfully expressed in Brevibacillus choshinensis hosts, and purified by a two-step column purification process. The antigen binding activity, structural characteristics, thermal stability, and stability in serum of these fusion proteins were examined. Results showed that the scFv with N-terminal fusing peptides proteins demonstrated increased LOX-1-binding activity without decrease in stability. These findings will help increase the application efficacy of LOX-1 targeting scFv in LOX-1-based therapy.http://dx.doi.org/10.1155/2017/8946935 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Wei Hu Qiuhong Xie Hongyu Xiang |
spellingShingle |
Wei Hu Qiuhong Xie Hongyu Xiang Improved scFv Anti-LOX-1 Binding Activity by Fusion with LOX-1-Binding Peptides BioMed Research International |
author_facet |
Wei Hu Qiuhong Xie Hongyu Xiang |
author_sort |
Wei Hu |
title |
Improved scFv Anti-LOX-1 Binding Activity by Fusion with LOX-1-Binding Peptides |
title_short |
Improved scFv Anti-LOX-1 Binding Activity by Fusion with LOX-1-Binding Peptides |
title_full |
Improved scFv Anti-LOX-1 Binding Activity by Fusion with LOX-1-Binding Peptides |
title_fullStr |
Improved scFv Anti-LOX-1 Binding Activity by Fusion with LOX-1-Binding Peptides |
title_full_unstemmed |
Improved scFv Anti-LOX-1 Binding Activity by Fusion with LOX-1-Binding Peptides |
title_sort |
improved scfv anti-lox-1 binding activity by fusion with lox-1-binding peptides |
publisher |
Hindawi Limited |
series |
BioMed Research International |
issn |
2314-6133 2314-6141 |
publishDate |
2017-01-01 |
description |
The oxidized low-density lipoprotein receptor-1 (LOX-1) targeted single-chain variable fragment (scFvs) is a promising molecule for the targeted delivery of imaging and therapeutic molecules of atherosclerotic diseases; however, its applications are limited by the inherent low antigen affinity. In this study, the three-dimensional (3D) model of the anti-LOX-1 scFv was constructed and its docking with the LOX-1 protein was developed. To improve the LOX-1-binding activity, the anti-LOX-1 scFv was designed to fuse with one of three LOX-1-binding heptapeptides, LTPATAI, FQTPPQL, and LSIPPKA, at its N-terminus and C-terminus and in the linker region, which have different LOX-1-binding interfaces with the anti-LOX-1 scFv analyzed by an array of computational approaches. These scFv/peptide fusions were constructed, successfully expressed in Brevibacillus choshinensis hosts, and purified by a two-step column purification process. The antigen binding activity, structural characteristics, thermal stability, and stability in serum of these fusion proteins were examined. Results showed that the scFv with N-terminal fusing peptides proteins demonstrated increased LOX-1-binding activity without decrease in stability. These findings will help increase the application efficacy of LOX-1 targeting scFv in LOX-1-based therapy. |
url |
http://dx.doi.org/10.1155/2017/8946935 |
work_keys_str_mv |
AT weihu improvedscfvantilox1bindingactivitybyfusionwithlox1bindingpeptides AT qiuhongxie improvedscfvantilox1bindingactivitybyfusionwithlox1bindingpeptides AT hongyuxiang improvedscfvantilox1bindingactivitybyfusionwithlox1bindingpeptides |
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1724759643582889984 |