Corresponding Author: Roland E. Kontermann, Institut für Zellbiologie und Immunologie, Universität Stuttgart, Allmandring 31, 70569 Stuttgart, Germany.
Mol Cancer Ther. 2014 Jan;13(1):101-11. doi: 10.1158/1535-7163.MCT-13-0396. Epub 2013 Oct 3.
We applied the immunoglobulin E (IgE) heavy-chain domain 2 (EHD2) as the covalently linked homodimerization module to generate antibody-scTRAIL fusion proteins. By fusing a humanized single-chain fragment variable (scFv) directed against EGFR to the N-terminus of the EHD2 and a single-chain derivative of TRAIL (scTRAIL) to the C-terminus of the EHD2, we produced a dimeric, tetravalent fusion protein. The fusion protein retained its binding activity for EGFR and TRAIL receptors. In vitro, the targeted antibody-scTRAIL fusion protein exhibited an approximately 8- to 18-fold increased cytotoxic activity compared with the untargeted EHD2-scTRAIL fusion protein. This resulted in increased antitumor activity in a subcutaneous Colo205 xenograft tumor murine model. In summary, the scFv-EHD2-scTRAIL fusion protein combines target cell selectivity with an increased TRAIL activity leading to improved antitumor activities.
我们应用免疫球蛋白 E（IgE）重链结构域 2（EHD2）作为共价连接的同二聚化模块，生成抗体-scTRAIL 融合蛋白。通过将针对 EGFR 的人源化单链片段可变区（scFv）融合到 EHD2 的 N 端，将 TRAIL 的单链衍生物（scTRAIL）融合到 EHD2 的 C 端，我们产生了二聚体、四价融合蛋白。融合蛋白保留了其对 EGFR 和 TRAIL 受体的结合活性。在体外，与非靶向 EHD2-scTRAIL 融合蛋白相比，靶向抗体-scTRAIL 融合蛋白的细胞毒性活性增加了约 8 至 18 倍。这导致在 Colo205 皮下异种移植肿瘤小鼠模型中抗肿瘤活性增加。总之，scFv-EHD2-scTRAIL 融合蛋白将靶细胞选择性与增加的 TRAIL 活性相结合，从而提高抗肿瘤活性。
