The Effect of Chemical Additives in Refolding of Recombinant Vascular Endothelial Growth Factor

Abstract

Background and Aim: The most important problem in the production of recombinant proteins in prokaryotic cells is the disruption of the function of these proteins due to their altered natural structure. The aim of present study is to identify the best chemicals dialysis buffer additives in order to improve the protein structure of recombinant Vascular Endothelial Growth Factor (VEGF) Methods & Materials: In this experimental study, different chemicals additives were selected using relevant software. After adding these additives to the recombinant VEGF dialysis buffer, their effect on the refolding of recombinant proteins and the differentiation of mesenchymal stem cells into endothelial cells was assessed by flow cytometry method. Ethical Considerations: This study obtained its ethical approval from the Research Ethics Committee of Arak University of Medical Sciences (Code: ARAKMU. REC.1394.199). Results: The results showed that the addition of arginine, cysteine and dithiothreitol (DTT) to dialysis buffer increases the differentiation of mesenchymal stem cells into endothelial cells. With the presence of sodium chloride (NaCl), cysteine, arginine and DTT in treated cells, the rate of specific Cluster Differentiation (CD) markers of endothelial cell (CD31/144) was at the highest level. Conclusion: Adding cysteine, arginine, DTT and NaCl to the dialysis buffer of recombinant VEGF had the greatest effect on the mesenchymal cell differentiation into endothelial cells.

Keywords

• cell differentiation
• vascular endothelial growth factor
• chemical additives
• cluster differentiation marker