High-yield expression, purification and characterization of tumor-targeted IFN-alpha2a

Cytotherapy. 2007;9(1):60-8. doi: 10.1080/14653240601094322.

Abstract

Background: IFN-alpha has been shown to be effective against hematologic malignancies. However, it is ineffective against most solid tumors and has not been satisfactory because of its toxicity.

Methods: The NGR (Asn-Gly-Arg) peptide is a tumor-homing peptide. In order to increase the anti-tumor activity of IFN-alpha2a and lower the dose, we coupled a cyclic NGR peptide with the C terminus of IFN-alpha2a (named IFN-alpha2a-NGR).

Results: The fusion protein was expressed in E. coli and purified by ion-exchange chromatography. The purity of IFN-alpha2a-NGR was >98% and the final purification yield of IFN-alpha2a-NGR was approximately 18 mg/L. The anti-tumor efficacy and the binding ability of IFN-alpha2a-NGR with tumor vasculature were investigated in vitro and in vivo.

Discussion: Our study has demonstrated that the anti-tumor efficacy of IFN-alpha2a-NGR is significantly increased in comparison with IFN-alpha2a, and IFN-alpha2a-NGR could selectively target tumor vessels. These data indicate that the tumor-homing peptide (NGR) can enhance the therapeutic efficacy of IFN-alpha2a against tumors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / isolation & purification
  • Antineoplastic Agents / pharmacology
  • Antineoplastic Agents / therapeutic use
  • Cell Line
  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Chromatography, High Pressure Liquid
  • Endothelial Cells / cytology
  • Endothelial Cells / drug effects
  • Endothelial Cells / physiology
  • Epithelial Cells / cytology
  • Epithelial Cells / drug effects
  • Epithelial Cells / virology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Humans
  • Interferon-alpha / genetics*
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • Microscopy, Confocal
  • Neoplasms / drug therapy*
  • Neoplasms / pathology
  • Oligopeptides / genetics*
  • Recombinant Fusion Proteins / isolation & purification*
  • Recombinant Fusion Proteins / pharmacology*
  • Recombinant Fusion Proteins / therapeutic use
  • Xenograft Model Antitumor Assays / methods

Substances

  • Antineoplastic Agents
  • Interferon-alpha
  • Oligopeptides
  • Recombinant Fusion Proteins
  • arginyl-glycyl-aspartic acid