【作者】 黄宇烽； 朱国华； 陈德宇；

【Author】 HUANG Yu-feng, ZHU Guo-hua, CHEN De-yu (Center of Medical Laboratory Science, Nanjing General Hospital of Nanjing Command ,PLA, Nanjing, 210002. Jiangsu, China)

【机构】 南京军区南京总医院检验中心；

【摘要】 目的在Sp2/0细胞中表达抗人L-PGDS人-鼠嵌合轻链。方法采用RT-PCR技术从稳定分泌抗人L-PGDS单抗的小鼠杂交瘤细胞中扩增抗体轻链可变区基因VL,并构建到人-鼠嵌合轻链表达载体pAG4622。重组质粒pAG4622/VL经DOTAP试剂转染入Sp2/0细胞,酶酚酸筛选后用ELISA和RT-PCR检测表达产物。结果在筛选后的转染细胞上清中检测到抗人L-PGDS人-鼠嵌合轻链蛋白,且其细胞中存在相应嵌合轻链mRNA。结论嵌合轻链的稳定表达,为后续抗人L-PGDS人-鼠嵌合抗体的获得奠定基础。更多还原

【Abstract】 Objective To express L-PGDS human-mouse chimeric light chain in Sp2/0 cells. Methods The variable region gene VL was cloned by RT-PCR from a mouse hybridoma cell line, which secreted McAb to human L-PGDS. The VL was then inserted into chimeric light chain expression vector pAG466 and expressed it in murine Sp2/0 cells. Results The transfected Sp2/0 clones selected by mycophenolic acid, expressed L-PGDS human-mouse chimeric light chain protein steadily, which was affirmed by ELISA and RT-PCR. Conclusion We expressed L-PGDS human-mouse chimeric light chain successfully, which laid a foundation to obtain its whole human-mouse chimeric antibodies and to research its therapeutic applications.更多还原