摘要
目的 在真核细胞中表达抗癌胚抗原 (carcinoembryonic antigen,CEA)人 -鼠嵌合抗体。方法 从分泌抗 CEA鼠单抗 C5 0的杂交瘤细胞中扩增、克隆可变区基因。测序鉴定后连入真核表达载体 ,导入二氢叶酸还原酶缺陷型中国仓鼠卵巢 (dihydrofolate reductase- deficient Chinese ham ster ovary,CHO- dhfr- )细胞进行表达。采用间接和竞争抑制 EL ISA检测表达产物的人源性和抗原结合特异性。结果 序列测定初步确定所克隆的是功能性抗体可变区基因。在转染细胞上清中测到抗 CEA嵌合抗体的表达。EL ISA试验证实该嵌合抗体具有人的恒定区 ,并与原鼠单抗 C5 0有相同的抗原结合特异性。结论 成功地在真核细胞中表达了抗 CEA人 -鼠嵌合抗体。
Objective To express a human mouse chimeric antibody against CEA antigen in eukaryotic cells.Methods The variable regions genes were amplified and cloned from the hybridoma that secrets anti CEA mouse monoclonal antibody C50.They were ligated into eukaryotic expression vectors after sequencing,and the were transfected into CHO dhfr cells for expression.The human constant regions and antigen specificity of expression products were identified by ELISA.Results The cloned antibody variable regions genes were approved functional by sequencing.The chimeric antibody ChC50 was detected in the culture supernatant of transfected CHO cells.The human constant regions and specificity of chimeric antibody to CEA were also detected the same as C50 by ELISA.Conclusion The anti CEA mouse human chimeric antibody was expressed successfully in eukaryotic cells.\;
出处
《实用肿瘤杂志》
CAS
北大核心
2001年第2期82-86,共5页
Journal of Practical Oncology
基金
国家九五攻关计划资助!(96 - 90 6 - 0 1- 0 2 1)
