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采用重组PCR技术获得抗多药耐药相关蛋白3(multidrug resistance protein 3,MRP3)的单链抗体(scFv)与人源可溶性肿瘤坏死因子相关凋亡诱导配体(soluble TNF-related apoptosis inducingligand,sTRAIL)的融合蛋白质的基因编码序列,利用原核表达载体pMAL-c2,构建含麦芽糖结合蛋白(maltose binding protein,MBP)标签肽的antiMRP3(scFv)-sTRAIL融合蛋白,经亲和层析柱纯化.获得纯化的antiMRP3(scFv)-sTRAIL融合蛋白,用MRP3阳性U251多形性胶质母细胞瘤做增殖抑制实验、细胞凋亡诱导实验,结果均显示具有明显的活性,而MBP无明显作用.上述结果表明,成功表达了antiMRP3(scFv)-sTRAIL融合蛋白,该融合蛋白具有诱导U251多形性胶质母细胞瘤细胞凋亡的活性,为开发靶向性抗肿瘤药物奠定了基础.
Single chain antibody (scFv) against multidrug resistance protein 3 (MRP3) and soluble TNF-related apoptosis inducing ligand (sTRAIL) were obtained by recombinant PCR. ), The antiMRP3 (scFv) -sTRAIL fusion protein containing the maltose binding protein (MBP) tag peptide was constructed by using prokaryotic expression vector pMAL-c2 and purified by affinity chromatography. Purified antiMRP3 (scFv) -sTRAIL fusion protein with MRP3 positive U251 glioblastoma proliferation inhibition experiments, apoptosis induction experiments, the results showed significant activity, MBP no significant effect. The above results The results showed that the fusion protein of antiMRP3 (scFv) -sTRAIL was successfully expressed, which has the activity of inducing apoptosis of U251 glioblastoma cells, which laid the foundation for the development of targeted anti-tumor drugs.