Recombinant expressing angiopep-2 fused anti-VEGF single chain Fab (scFab) could cross blood–brain barrier and target glioma
Abstract In 2009, the FDA approved bevacizumab for the treatment of adult patients diagnosed with recurrent glioblastoma. However, the poor permeability of the macromolecules across the blood–brain barrier, determined by multifactorial anatomical and physiological milieu, restricts the clinical ther...
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doaj-a9be7bb6c90a47a287ee4b57302bcebe2020-11-25T03:41:36ZengSpringerOpenAMB Express2191-08552019-10-019111210.1186/s13568-019-0869-3Recombinant expressing angiopep-2 fused anti-VEGF single chain Fab (scFab) could cross blood–brain barrier and target gliomaXuemei Ji0Hongyan Wang1Yue Chen2Junfei Zhou3Yu Liu4State Key Laboratory of Natural Medicines, School of Life Science and Technology, China Pharmaceutical UniversityState Key Laboratory of Natural Medicines, School of Life Science and Technology, China Pharmaceutical UniversityState Key Laboratory of Natural Medicines, School of Life Science and Technology, China Pharmaceutical UniversityState Key Laboratory of Natural Medicines, School of Life Science and Technology, China Pharmaceutical UniversityState Key Laboratory of Natural Medicines, School of Life Science and Technology, China Pharmaceutical UniversityAbstract In 2009, the FDA approved bevacizumab for the treatment of adult patients diagnosed with recurrent glioblastoma. However, the poor permeability of the macromolecules across the blood–brain barrier, determined by multifactorial anatomical and physiological milieu, restricts the clinical therapeutic effect of bevacizumab. The low-density lipoprotein receptor related protein 1 (LRP1) is highly expressed in the endothelial cells of the brain capillary and the glioma cells. Angiopep-2 (ANG) is a 19-aa oligopeptide that can bind to LRP1 and penetrate the blood–brain barrier by receptor-mediated transport. Therefore, ANG can be used as a dual-targeting drug delivery carrier into the brain and the glioma sites. In this study, ANG gene was fused with the C-terminal domain of single-chain antigen binding fragment (scFab) of the anti-VEGF antibody and recombinant scFab-ANG protein was expressed and purified using Rosatte (DE3) strain. We confirmed that ANG could carry anti-VEGF-scFab, penetrate a three-dimensional model of the brain tumor, and cross the hCMEC/D3 monolayer in the in vitro blood–brain barrier model. The animal experiments demonstrated that 3 h after the tail intravenous protein injection, the fluorescent signals in the brains of the mice in the scFab-ANG group were stronger than that in the scFab group. Furthermore, the study of the in situ rat glioma model shows that scFab-ANG could target glioma while anti-VEGF-scFab could not. These findings indicate that scFab-ANG had stronger transepithelial permeability and glioma targeting capacity. Thus, it can be a potential candidate drug for glioblastoma therapy.http://link.springer.com/article/10.1186/s13568-019-0869-3GliomaBlood–brain barrierAngiopep-2ScFab-ANGTransepithelial permeability |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Xuemei Ji Hongyan Wang Yue Chen Junfei Zhou Yu Liu |
spellingShingle |
Xuemei Ji Hongyan Wang Yue Chen Junfei Zhou Yu Liu Recombinant expressing angiopep-2 fused anti-VEGF single chain Fab (scFab) could cross blood–brain barrier and target glioma AMB Express Glioma Blood–brain barrier Angiopep-2 ScFab-ANG Transepithelial permeability |
author_facet |
Xuemei Ji Hongyan Wang Yue Chen Junfei Zhou Yu Liu |
author_sort |
Xuemei Ji |
title |
Recombinant expressing angiopep-2 fused anti-VEGF single chain Fab (scFab) could cross blood–brain barrier and target glioma |
title_short |
Recombinant expressing angiopep-2 fused anti-VEGF single chain Fab (scFab) could cross blood–brain barrier and target glioma |
title_full |
Recombinant expressing angiopep-2 fused anti-VEGF single chain Fab (scFab) could cross blood–brain barrier and target glioma |
title_fullStr |
Recombinant expressing angiopep-2 fused anti-VEGF single chain Fab (scFab) could cross blood–brain barrier and target glioma |
title_full_unstemmed |
Recombinant expressing angiopep-2 fused anti-VEGF single chain Fab (scFab) could cross blood–brain barrier and target glioma |
title_sort |
recombinant expressing angiopep-2 fused anti-vegf single chain fab (scfab) could cross blood–brain barrier and target glioma |
publisher |
SpringerOpen |
series |
AMB Express |
issn |
2191-0855 |
publishDate |
2019-10-01 |
description |
Abstract In 2009, the FDA approved bevacizumab for the treatment of adult patients diagnosed with recurrent glioblastoma. However, the poor permeability of the macromolecules across the blood–brain barrier, determined by multifactorial anatomical and physiological milieu, restricts the clinical therapeutic effect of bevacizumab. The low-density lipoprotein receptor related protein 1 (LRP1) is highly expressed in the endothelial cells of the brain capillary and the glioma cells. Angiopep-2 (ANG) is a 19-aa oligopeptide that can bind to LRP1 and penetrate the blood–brain barrier by receptor-mediated transport. Therefore, ANG can be used as a dual-targeting drug delivery carrier into the brain and the glioma sites. In this study, ANG gene was fused with the C-terminal domain of single-chain antigen binding fragment (scFab) of the anti-VEGF antibody and recombinant scFab-ANG protein was expressed and purified using Rosatte (DE3) strain. We confirmed that ANG could carry anti-VEGF-scFab, penetrate a three-dimensional model of the brain tumor, and cross the hCMEC/D3 monolayer in the in vitro blood–brain barrier model. The animal experiments demonstrated that 3 h after the tail intravenous protein injection, the fluorescent signals in the brains of the mice in the scFab-ANG group were stronger than that in the scFab group. Furthermore, the study of the in situ rat glioma model shows that scFab-ANG could target glioma while anti-VEGF-scFab could not. These findings indicate that scFab-ANG had stronger transepithelial permeability and glioma targeting capacity. Thus, it can be a potential candidate drug for glioblastoma therapy. |
topic |
Glioma Blood–brain barrier Angiopep-2 ScFab-ANG Transepithelial permeability |
url |
http://link.springer.com/article/10.1186/s13568-019-0869-3 |
work_keys_str_mv |
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