[单克隆抗体F(ab')2片段介导的阿霉素免疫纳米球的制备及其对肝癌细胞的体外杀伤作用]

[Preparation and in vitro killing effect of adriamycin-loaded immunonanosphere against hepatoma led by F (ab')2 Fragment of monoclonal antibodies].

作者信息

Li Yunchun, Cai Meiying, Wang Zhongqiong, Liu Xiaobo, Liang Zhenglu

机构信息

Department of Nuclear Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.

出版信息

Hua Xi Yi Ke Da Xue Xue Bao. 2002 Jan;33(1):8-11, 14.

PMID:12599415

Abstract

OBJECTIVE

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.

METHODS

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.

RESULTS

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.

CONCLUSION

HAb18 F(ab')2-ADR-HSA-NP has a good character for in vitro specific targeting to bind and kill human hepatoma cell.

摘要

目的

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。

方法

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。

结果

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。