Expression, purification, and characterization of a neovasculature targeted rmhTNF-alpha in Escherichia coli.
The tumor vasculature is a suitable target for cancer treatment. RGD-4C (CDCRGDCFC) peptide can bind to human alphav 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-alpha mutant (rmhTNF-alpha) that we previously constructed has been proved to have stronger antitumor effect compared with TNF-alpha. To increase antitumor effect and lower toxicity of rmhTNF-alpha, we coupled RGD4C to the N-terminal of rmhTNF-alpha (termed RGD4C-rmhTNF) and expressed RGD4C-rmhTNF in Escherichia coli. Here, we describe the expression, purification, and characterization of RGD4C-rmhTNF.
Wang
Hui
H
Biotechnology Center of The Fourth Military Medical University, 17 Changle West Road, 710032 Xi'an, PR China.
Yan
Zhen
Z
Shi
Jihong
J
Han
Wei
W
Zhang
Yingqi
Y
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects; D039302:Integrin alphaVbeta3 / Q000737:chemistry / Q000187:drug effects; D051379:Mice; D009154:Mutation; D009389:Neovascularization, Pathologic / Q000235:genetics*; D009842:Oligopeptides / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology
D039302:Integrin alphaVbeta3; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha