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

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 BL۲۱ (DE۳) competent cells were transformed with pET۳۲a-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 ۰.۸ mg/۱۰۰ w/w) was used for external wound (۲۵×۱۵mm 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 ۴۵ kilodaltons (kDa) and concentration of ۰.۸ 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 pET۳۲a in E. coli, possesses acceptable structure and has wound healing capability.

Keywords:

Biological activity , Recombinant , VEGF-A , Wound healing