Li Yunchun, Cai Meiying, Wang Zhongqiong, Liu Xiaobo, Liang Zhenglu
Department of Nuclear Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
Hua Xi Yi Ke Da Xue Xue Bao. 2002 Jan;33(1):8-11, 14.
To study the preparation method and in vitro killing effect of adriamycin (ADR)-loaded human serum albumin (HSA) immunonanosphere (HAb18 F(ab')2-ADR-HSA-NP) against hepatoma led by F(ab')2 fragment of human hepatoma specific monoclonal antibody HAb18.
After ADR loaded HSA nanosphere (ADR-HSA-NP) was prepared in the emulsifying high temperature solidifying way, HAb18 F(ab')2-ADR-HSA-NP was prepared using the modified N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP) method. In vitro binding characters of HAb18 F(ab')2-ADR-HSA-NP and ADR-HSA-NP and hepatoma cell SMMC-7721 were observed under optical microscopy and electronic microscopy. In vitro effects of killing hepatoma cell SMMC-7721 of two microspheres were determined using the method of 3H-TdR.
The surfaces of HAb18 F(ab')2-ADR-HSA-NP gave out bright yellow-green fluorescence after it was dyed with fluorescent agent, whereas ADR-HSA-NP did not give out fluorescence. HAb18 F(ab')2-ADR-HSA-NP could integrate with hepatoma cell SMMC-7721 and effectively killed hepatoma cell SMMC-7721 with dose dependence, but ADR-HSA-NP could not obviously integrate and kill SMMC-7721. Neither of the two microspheres could bind and kill human large intestine cancer cell SW1116.
HAb18 F(ab')2-ADR-HSA-NP has a good character for in vitro specific targeting to bind and kill human hepatoma cell.
研究以人肝癌特异性单克隆抗体HAb18的F(ab')2片段为导向的阿霉素（ADR）负载人血清白蛋白（HSA）免疫纳米球（HAb18 F(ab')2-ADR-HSA-NP）的制备方法及其对肝癌细胞的体外杀伤作用。
采用乳化高温固化法制备ADR负载HSA纳米球（ADR-HSA-NP）后，用改良的N-琥珀酰亚胺基-3-(2-吡啶二硫代)丙酸酯（SPDP）法制备HAb18 F(ab')2-ADR-HSA-NP。在光学显微镜和电子显微镜下观察HAb18 F(ab')2-ADR-HSA-NP和ADR-HSA-NP与肝癌细胞SMMC-7721的体外结合特性。采用3H-TdR法测定两种微球对肝癌细胞SMMC-7721的体外杀伤作用。
用荧光剂染色后，HAb18 F(ab')2-ADR-HSA-NP表面发出亮黄绿色荧光，而ADR-HSA-NP不发出荧光。HAb18 F(ab')2-ADR-HSA-NP能与肝癌细胞SMMC-7721结合，并呈剂量依赖性有效地杀伤肝癌细胞SMMC-7721，但ADR-HSA-NP不能明显结合和杀伤SMMC-7721。两种微球均不能结合和杀伤人大肠癌细胞SW1116。
HAb18 F(ab')2-ADR-HSA-NP具有良好的体外特异性靶向结合和杀伤肝癌细胞的特性。
