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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。
Objective To express a human chimeric gene engineered antibody against human lung cancer monoclonal antibody (mAb) LC1. Methods The VL and VH genes of mAbLC 1 were inserted into the plasmid pWS1 to construct a plasmid pLC 1 expressing the mAbLC 1 human chimeric Fab fragment gene and transformed into E. coli TG 1 and expressed therein. RESULTS: Enzyme digestion analysis showed that the inserted VL and VH genes of mAbLC 1 were in the correct orientation. This vector could express the expected molecular weight of the protein in E. coli TG1, and the expression product had a strong competition for antigen activity with the mAb. Conclusions The engineered antibodies of human and mouse chimeric anti-human lung cancers with higher affinity activity were successfully expressed.