胶质瘤基因工程单链抗体的制备及其抗瘤效应的初步研究

Preparation of single chain antibody by gene engineering and preliminary study on its anti - tumor activity against glioma

目的 构建并高效表达抗胶质瘤单链抗体（ScFv）,为基因工程双功能抗体及免疫毒素的制备奠定基础。方法 以抗胶质瘤杂交瘤细胞重、轻链可变区基因构建ScFv基因,并克隆人pET-20b表达载体,在大肠杆菌BL21 DE3中诱导表达,以Western印迹及酶联免疫吸附测定（ELISA）检测表达产物。结果 测序证实基因构建正确,表达产物相对分子质量（Mr）为28000,表达量占菌体总蛋白的15.0％,且保留了胶质瘤相关抗原的特异结合活性。结论 初步得到有活性的胶质瘤ScFv,为脑肿瘤的分子导向诊治提供了新的选择。

Objective To construct the anti -glioma ScFv gene and express it in E. coli with high level. Methods The VH and VL gene were combined with a (Gly4Ser) 3 linker to construct ScFv gene against glioma, and then cloned into the expression vector pET -20b, and expressed in E. coli BL21 DE3. The activities of ScFv were assayed by Western blot and enzyme - linked immunosorbent assay ( ELISA). Results DNA sequencing proved that ScFv gene was cloned correctly into the expression vector. The relative molecular mass of the expressed product was 28 000. The expressed product amounted to 15. 0% of the total bacterial protein, and retained its anti - tumor activities. Conclusion The preparation of anti - glioma ScFv may offer new choices for molecular targeting therapy against glioma.