单抗F(ab′)_2段导向抗肝癌阿霉素免疫毫微球的制备及其体外杀伤癌细胞作用  被引量:6

Preparation and in Vitro Killing Effect of Adriamycin-loaded Immunonanosphere Against Hepatoma Led by F(ab′)_2 Fragment of Monoclonal Antibodies

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作  者:李云春[1] 蔡美英[2] 王仲琼 刘晓波 梁正路[1] 

机构地区:[1]四川大学华西医院核医学科,成都610041 [2]华西基础医学与法医学院免疫学教研室 [3]华西医院检验科

出  处:《华西医科大学学报》2002年第1期8-11,14,共5页Journal of West China University of Medical Sciences

基  金:卫生部科研基金资助 (编号 98-1-2 2 5 )

摘  要:目的 制备人肝癌特异性单克隆抗体 HAb18的 F(ab′) 2 片段导向抗肝癌阿霉素 (ADR)白蛋白(HSA)免疫毫微球 (HAb18F(ab′) 2 - ADR- HSA- NP) ,并研究其体外靶向肝癌细胞和杀伤肝癌细胞的作用。方法 采用乳化高温固化法制备阿霉素白蛋白毫微球 (ADR- HSA- NP)后 ,应用改进的 N-琥珀酰亚胺基 - 3- (2 -吡啶二硫 )丙酸酯 (SPDP)法制备 HAb18F(ab′) 2 - ADR- HSA- NP。在光镜和电镜下观察体外 HAb18F(ab′) 2 - ADR- HSA- NP和 ADR- HSA- NP与肝癌细胞株 SMMC- 772 1的结合特性 ,并用 3H- Td R法测定两种微球体外杀伤肝癌细胞株SMMC- 772 1的作用。结果 在荧光染色后 ,HAb18F(ab′) 2 - ADR- HSA - NP表面发出明亮黄绿色荧光 ,而 ADR-HSA- NP未见荧光。 HAb18F(ab′) 2 - ADR- HSA- NP能结合肝癌细胞株 SMMC- 772 1,且能呈剂量依赖地有效杀伤肝癌细胞株 SMMC- 772 1,而 ADR- HSA- NP不能结合和明显杀伤肝癌细胞株 SMMC- 772 1。两种微球均不能结合和杀伤人大肠癌细胞株 SW1116。结论  HAb18F(ab′) 2 - ADR- HSA -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.

关 键 词:单抗片段 免疫毫微球 肝癌细胞 杀伤活性 单克隆抗体 阿霉素 

分 类 号:R735.7[医药卫生—肿瘤] R730.51[医药卫生—临床医学]

 

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