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目的在哺乳动物细胞内表达抗体细胞因子融合蛋白183B2scFv Interleukin2,为卵巢癌提供一种新的治疗方法。方法通过基因工程的方法将两段基因IL2和183B2scFv开放读框的编码序列克隆在一起,在CHO K1细胞内人巨细胞病毒启动子的作用下表达可溶性融合蛋白,并检测其对肿瘤细胞的靶向性。结果采用哺乳动物细胞表达,系统表达的这种小分子融合蛋白,既保留了与卵巢癌OC183B2抗原结合的特性,又保持了IL2的生物学活性。融合蛋白在细胞培养上清中保持稳定,更重要的是融合蛋白将IL2靶向表达OC183B2抗原的卵巢癌细胞表面化的同时,又能刺激IL2依赖细胞株的增殖,从而诱发局部有效的抗肿瘤反应。既提高了融合蛋白的穿透组织能力和肿瘤组织部位的浓聚,又避免了大剂量全身应用细胞因子引起的副作用。结论真核表达系统表达的融合蛋白具有很好的生物学活性。
Objective To express antibody cytokine fusion protein 183B2scFv Interleukin2 in mammalian cells to provide a new therapeutic method for ovarian cancer. Methods The coding sequence of two open reading frames of IL2 and 183B2 scFv genes were cloned by genetic engineering. The soluble fusion protein was expressed under the action of human cytomegalovirus promoter in CHO K1 cells and its target on tumor cells Sexuality. Results The small molecule fusion protein expressed in mammalian cells and expressed in the system retained the characteristic of binding to OC183B2 antigen of ovarian cancer and maintained the biological activity of IL2. The fusion protein remains stable in cell culture supernatants. More importantly, the fusion protein targets IL2 on the surface of ovarian cancer cells expressing the OC183B2 antigen while stimulating the proliferation of IL2-dependent cell lines to induce locally effective antitumor responses . It not only improves the penetrating ability of the fusion protein and the accumulation of tumor tissue, but also avoids the side effects caused by high-dose systemic application of cytokines. Conclusion The fusion protein expressed by eukaryotic expression system has good biological activity.