Expression, purification, and characterization of a neovasculature targeted rmhTNF-α in Escherichia coli

The tumor vasculature is a suitable target for cancer treatment. RGD-4C (CDCRGDCFC) peptide can bind to human αv integrins, which are known to be selectively expressed in human tumor blood vessels. Some studies showed that coupling anticancer drugs or peptides to the RGD peptides yielded compounds with increased efficacy against tumors and lowered toxicity to normal tissues in mice. TNF-α mutant (rmhTNF-α) that we previously constructed has been proved to have stronger antitumor effect compared with TNF-α. To increase antitumor effect and lower toxicity of rmhTNF-α, we coupled RGD4C to the N-terminal of rmhTNF-α (termed RGD4C-rmhTNF) and expressed RGD4C-rmhTNF in Escherichia coli. Here, we describe the expression, purification, and characterization of RGD4C-rmhTNF.