PUBLICATION
Expression, purification and characterization of a vascular endothelial growth factor fusion protein
- Authors
- Zhang, Y., Tong, Y., Gao, M., Luo, C., Song, X., Gao, X., Yao, W.
- ID
- ZDB-PUB-160316-3
- Date
- 2016
- Source
- Biotechnology Letters 38(7): 1115-20 (Journal)
- Registered Authors
- Zhang, Yu
- Keywords
- Angiogenesis, Expression and purification, Vascular endothelial growth factor B, Zebrafish
- MeSH Terms
-
- Zebrafish
- Recombinant Proteins/genetics
- Recombinant Proteins/metabolism*
- Recombinant Proteins/pharmacology
- Humans
- PubMed
- 26976430 Full text @ Biotechnol. Lett.
Abstract
Objectives To prepare recombinant tPep-(vascular endothelial growth factor) VEGF-B and assess its biological activity.
Results This new VEGF fusion protein was constructed using a targeting peptide and prepared using E.coli. The tPep-VEGF-B was refolded from inclusion bodies and purified using affinity chromatography. Its bioactivity was determined in vitro using proliferation assay and wounding healing assay, and in vivo in zebrafish. By using the optimized downstream process, recombinant tPep-VEGF-B can be obtained with a purity of >90 % and a yield of 80 mg protein/l culture medium. The refolded protein is highly effective in promoting cell migration in vitro and in enhancing angiogenesis in vivo.
Conclusion We have constructed a new VEGF fusion protein with potential therapeutic application in treating metabolic diseases.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping