Introduction
Platelet-derived growth factor (PDGF) is a potent mitogen and chemoattractant for many cells of mesenchymal origin, such as fibroblasts, activated macrophages, and smooth muscle cells. PDGF plays critical roles in angiogenesis, embryogenesis, inflammation, and cell differentiation; therefore, it can be beneficial in wound healing. It has three dimer forms: PDGF-AA, AB, and BB, produced by platelets, activated macrophages, activated fibroblasts, keratinocytes, and some other cells in the body. The therapeutic potential of recombinant human PDGF-BB in wound healing and diabetic ulcers has been proven. The objective of this study was to produce recombinant human PDGF-BB protein in E. coli and its puri... More
Introduction
Platelet-derived growth factor (PDGF) is a potent mitogen and chemoattractant for many cells of mesenchymal origin, such as fibroblasts, activated macrophages, and smooth muscle cells. PDGF plays critical roles in angiogenesis, embryogenesis, inflammation, and cell differentiation; therefore, it can be beneficial in wound healing. It has three dimer forms: PDGF-AA, AB, and BB, produced by platelets, activated macrophages, activated fibroblasts, keratinocytes, and some other cells in the body. The therapeutic potential of recombinant human PDGF-BB in wound healing and diabetic ulcers has been proven. The objective of this study was to produce recombinant human PDGF-BB protein in E. coli and its purification.
Methods
In this research study, the human PDGF-B gene sequence was obtained and synthesized after optimization. Using a prokaryotic expression vector, pET-15b, pET-15b-PDGF-B was constructed. The expression and purification of the recombinant human PDGF-B in a prokaryotic host, Escherichia coli BL21 (DE3), under T7 promoter was performed. Protein purification was done on Ni2+ affinity chromatography (NTA) column.
Results
The recombinant human PDGF-BB protein, as a potential therapeutic target, was produced by cloning and protein expression of human PDGF-B gene in E. coli, then was purified. The purified recombinant human protein, pET-PDGF-B, revealed a sharp band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with a molecular weight of about 12 kDa.
Conclusion
The recombinant construction of prokaryotic expression vector, pET15b-PDGF-B, and the production of PDGF-BB protein provide a foundation for further studies about the function of PDGF-BB and production of biologically active PDGF-BB protein.
