抗人肺癌mAbLC-1的人-鼠嵌合Fab片段基因表达质粒的构建与表达

Construction and expression of recombinant plasmid pLC-1 bearing human mouse chimeric Fab fragment gene of mAb LC-1 to human pulmonary carcinoma

目的表达抗人肺癌单克隆抗体(mAb)LC 1的人 鼠嵌合基因工程抗体。方法将mAbLC 1的VL 和VH 基因插入质粒 pWS1 中 ,构建成表达mAbLC 1人 鼠嵌合Fab片段基因的质粒pLC 1 ,转化大肠杆菌TG 1并在其中表达。结果酶切鉴定表明 ,mAbLC 1的VL 和VH 基因插入方向正确 ,该载体能在大肠杆菌TG1中表达预期相对分子质量的蛋白 ,表达产物对mAb具有较强的竞争结合抗原的活性。结论成功地表达了具有较高亲和活性的人 鼠嵌合抗人肺癌的基因工程抗体。

Aim To express human mouse chimeric genetic engineering antibody of monoclonal antibody(mAb)LC 1 to human pulmonary carcinoma. Methods VL and VH genes of the mAb LC 1 were cloned into plasmid pSW1 and then plasmid pLC 1 expressing human mouse chimeric Fab fragment gene of the mAb LC 1 was constructed. The plasmid pLC 1 was transformed into E.coli TG 1 and expressed in it. Results Restriction enzyme digestion showed that the VL and VH genes of the mAb LC 1 were inserted into plasmid pSW1 in correct direction. Expression products of plasmid pSW1 in E.coli TG 1 could compete against mAb LC 1 in binding to antigen. Conclusion Human mouse chimeric genetic engineering antibody to human pulmonary carcinoma is successfully expressed, with higher affinity activity.