Construction and expression of recombinant plasmid pLC-1 bearing human mouse chimeric Fab fragment gene of mAb LC-1 to human pulmonary carcinoma
CHEN Liang, XIE He huang, WANG Jue, FAN Zhao juan, LIN Xin kai, CHEN Song hua, LIU Wei, GE Xi ruiInstitute of Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China
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.