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

目的观察抗肺腺癌单链抗体(ScFv)2A7-1-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2A7-1E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2A7-1、ADM制备结合物,分光光度计测定结合物中所含2A7-1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2A7-1-ADM结合物中,ADM和2A7-1的克分子比为3：1。连接后的2A7-1-ADM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2A7-1-ADM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。

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

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 adenocarcinoma 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.