Rational design of an EGF-IL18 fusion protein: implication for developing tumor therapeutics.

Abstract

Interleukin-18 (IL-18) is a proinflammatory cytokine. This protein has a role in regulating immune responses and exhibits significant anti-tumor activities. Epidermal growth factor (EGF) is an important growth factor that plays a central role in the regulation of cell cycle and differentiation. It was proposed that a targeted delivery of IL-18 by generation of IL-18-EGF fusion protein might decrease adverse effects and result in enhancing cytotoxic and antitumor activities. In the present study, a fusion protein, consisting of EGFR binding domain fused to human IL-18 mature peptide via a linker peptide of (Gly(4)Ser) 3, was constructed and expressed in the insect cell line Sf9 using Bac-to-Bac baculovirus expression system. We showed that the purified recombinant fusion protein induced similar levels of IFN-gamma to that of native IL-18 protein in human PBMC in the presence of ConA. Furthermore, EGF receptor competitive test in human epithelial cancer A431 cell line showed that EGF-IL18 fusion protein can specifically bind with EGFR by competing with native EGF protein. These suggest that this rationally designed protein can be further developed as novel tumor therapeutics.

• Authors: Jian-Xin Lu, Ying Peng, Zhe-Feng Meng, Li-Qin Jin, Yong-Sui Lu, Min-Xin Guan
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 334 Issue 1 Pg. 157-61 (Aug 19 2005) ISSN: 0006-291X [Print] United States
• PMID: 15993840 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Interleukin-18
• Recombinant Fusion Proteins
• Epidermal Growth Factor
• ErbB Receptors

Topics

• Animals
• Carcinoma, Squamous Cell (genetics, metabolism, therapy)
• Cell Line
• Drug Delivery Systems (methods)
• Drug Design
• Epidermal Growth Factor (genetics, metabolism, therapeutic use)
• ErbB Receptors (metabolism)
• Gene Targeting (methods)
• Humans
• Interleukin-18 (genetics, metabolism, therapeutic use)
• Protein Engineering (methods)
• Recombinant Fusion Proteins (metabolism)
• Spodoptera (genetics, metabolism)
• Transfection (methods)