Heterologous expression of a truncated form of human recombinant vascular endothelial growth factor-A and its biological activity in wound healing

Autor: Mohsen Khaki, Ali Hatef Salmanian, Ghasem Mosayebi, Maryam Baazm, Saeed Babaei, Neda Molaee, Hamid Abtahi
Jazyk: angličtina
Rok vydání: 2017
Předmět:
Zdroj: Iranian Journal of Basic Medical Sciences, Vol 20, Iss 7, Pp 791-797 (2017)
Druh dokumentu: article
ISSN: 2008-3866
2008-3874
DOI: 10.22038/ijbms.2017.9011
Popis: Objective(s): Vascular endothelial growth factor (VEGF) is one of the most effective proteins in angiogenesis, mesenchymal stem cells (MSCs) differentiation and wound healing. These abilities are therapeutic potential of VEGF in diabetic retinopathy, nephropathy and other tissue damage circumstances. In this study, recombinant VEGF was produced in Escherichia coli (E. coli) system and then biological activity of this protein was evaluated in animal wound healing. Materials and Methods: E. coli BL21 (DE3) competent cells were transformed with pET32a-VEGF clone and induced by isopropyl-β-D-thio-galactoside (IPTG). The recombinant protein was purified byaffinity chromatography. Recombinant VEGF-A-based ointment (VEGF/Vaseline 0.8 mg/100 w/w) was used for external wound (25×15mm thickness) healing in animal model. In vivo activity of ointment was evaluated by clinical evidences and cytological microscopic assessment. Results: The recombinant protein with molecular weight of 45 kilodaltons (kDa) and concentration of 0.8 mg/ml was produced.Immunoblotting data showed that the antigenic region of VEGF can be expressed in E. coli and the recombinant protein has similar epitopes with close antigenic properties to the natural form. Macroscopic findings and microscopic data showed that the recombinant VEGF-A ointment was effective on excisional wound healing. Conclusion: Recombinant VEGF-A produced by pET32a in E. coli, possesses acceptable structure and has wound healing capability.
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