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Rational design of an EGF-IL18 fusion protein: implication for developing tumor therapeutics
Authors:Lu Jian-Xin  Peng Ying  Meng Zhe-Feng  Jin Li-Qin  Lu Yong-Sui  Guan Min-Xin
Institution:Institute of Cellular and Molecular Medicine, School of Laboratory Medicine, Wenzhou Medical College, Wenzhou, Zhejiang 325035, China.
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.
Keywords:EGF-IL18 fusion protein  Tumor therapeutics  Cytokine  Expression  Targeting  Anti-tumor  IL-18  EGFR binding domain  Fusion proteins  Baculovirus expression system  Targeting tumor cells
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