单链抗体阿霉素结合物对人肺腺癌细胞体外生长的抑制作用  

Growth inhibitory effect of adriamycin conjugated to single-chain antibody on human lung adenocarcinoma in vitro

作　　者：岳文涛[1] 赖百塘[1] 汪蕙[1] 湛秀萍[1] 王玥[1] 

机构地区：[1]北京市结核病胸部肿瘤研究所细胞室,101149

出　　处：《中华肿瘤杂志》2004年第12期718-721,共4页Chinese Journal of Oncology

摘　　要：目的观察抗肺腺癌单链抗体(ScFv)2A71阿霉素(ADM)结合物对人肺腺癌细胞体外生长的抑制作用。方法从抗肺癌可溶性ScFv抗体分泌克隆2A71上清提取可溶性抗体,浓缩、亲和层析纯化,以分光光度计测定浓度。戊二醛法连接2A71、ADM制备结合物,分光光度计测定结合物中所含2A71和ADM的浓度,计算结合比例。集落形成试验观察该结合物对肺癌细胞体外生长的抑制作用。结果制备的2A71ADM结合物中,ADM和2A71的克分子比为31。连接后的2A71ADM免疫结合物仍可特异结合肺腺癌A2细胞,其对肺癌细胞的抑制作用是单纯使用ADM的4倍。结论2A71ADM免疫结合物对肺癌细胞的生长抑制作用明显强于单独使用ADM,具有潜在的临床应用价值。Objective To evaluate the growth inhibitory effect of adriamycin (ADM) conjugated to an anti-lung cancer single-chain antibody (ScFv) 2A7-1 on lung adenocarcinoma cell line A2 in vitro. Methods 2A7-1 cell culture medium was concentrated by ultra-filtration (with Amicon P10Z filter), and soluble ScFv was purified using RPAS purification kit. ADM was conjugated to 2A7-1 by glutaraldehyde. A_~280 and A_~490 of the conjugate 2A7-1-ADM were determined by spectrophotometry and the molar ratio of 2A7-1 to ADM was calculated. Immunoreactivity of the conjugate was detected by immunohistochemistry. Its growth inhibitory effect on lung adenocarcioma cell line A2 was determined by colony formation assay in vitro. Results The molar ratio of 2A7-1 to ADM was 1∶3.2. The conjugate strongly reacted with A2 cell. Its growth inhibitory effect on A2 cells was 4 times as potent as ADM. Conclusion Adriamycin conjugated to anti-lung cancer single-chain antibody 2A7-1 has much higher cytotoxic activity than unconjugated adriamycin against human lung adenocarcinoma.

关 键 词：单链抗体阿霉素结合物 肺腺癌 癌细胞 体外生长 抑制作用 

分 类 号：R734.2[医药卫生—肿瘤]