Browsing by Subject "Bioassay"

Vascular endothelial growth factor C (VEGF-C) is a key lymphangiogenic protein and potential therapeutic target, but its production in bioactive form from bacteria has been challenging. This study focused on the purification method of VEGF-C from Escherichia coli (E. coli) using three-step chromatographic approaches involving anion exchange, hydrophobic interaction, and size exclusion chromatography. While the major purified species migrated at the 35 kDa size, it lacked biological activity in a Ba/F-3-VEGFR-3 bioassay, likely due to interference from the partially cleaved maltose binding protein fusion tag. The maximum yield of 0.311 mg of VEGF-C per liter of bacterial culture represented a modest 3.11-fold increase over previous Ni-affinity chromatography but a 1.93-fold decrease compared to amylose affinity chromatography purification. Further work is needed to optimize tag removal and purification to enable larger-scale production of bioactive VEGF-C for research and potential therapeutic applications. While this study demonstrates conventional chromatography can purify VEGF-C, but highlights challenges in obtaining high yields of the bioactive VEGF-C.