摘要
构建人肝癌特异的免疫毫微粒 ,探讨免疫毫微粒体外对靶细胞的特异性结合特性及杀伤活性。方法 :采用异型双功能交联剂SPDP将人肝癌单抗HAb18与载米托蒽醌的白蛋白毫微粒化学偶联 ,构建人肝癌特异的免疫毫微粒 ;采用玻片凝集试验 ,免疫荧光法及荧光染色阻断法 ,花环形成实验及花环形成阻断实验 ,扫描电镜 ,3H TdR掺入试验证明抗体与载药毫微粒偶联及免疫毫微粒能特异性结合并杀伤靶细胞SMMC 772 1人肝癌株。结果 :HAb18抗体已偶联到载药毫微粒上 ;免疫毫微粒能良好地与靶细胞特异性结合 ,对靶细胞具有剂量依赖性、选择性杀伤作用。结论 :SPDP偶联方法能用于构建免疫毫微粒 ;人肝癌特异免疫毫微粒体外能特异性结合并杀伤人肝癌细胞株。
Objective:To construct human hepatoma-specific immunonanoparticles and observe in vitro binding and killing of target cells by the immunonanoparticles.Methods:Hetero bifunctional crosslinker SPDP was used to couple covalently McAb HAb18(anti-human hepatoma) with mitoxantrone-loaded bovine serum albumin nanoparticles(DHAQ-BSA-NP). A lot of in vitro experiments,such as slide agglutination test,immunofluorescent assay,immunofluorescent blocking test,rosset formation test,rosset formation blocking test and scanning electron microscopy etc,were used to confirm coupling of antibodies to nanoparticles and specific binding of immunonanoparticles with target cells. 3 H-TdR incorporation test was used to assay cytotoxicity of immunonanoparticles to target cells.Results:McAb HAb18 was coupled to DHAQ-BSA-NP. The immunonanoparticles could bind specifically to human hepatoma SMMC-7721 cells and exert selective killing effects on the target cells with dose-dependence.Conclusion:Conjugation technique via SPDP is a good method for constructing immunonanoparticles. Human hepatoma-specific-immunonanoparticles have capacity of in vitro selective coating and killing human hepatoma cell line.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2000年第5期262-265,共4页
Chinese Journal of Immunology
基金
卫生部科学研究基金资助项目 !(No .98 1 2 2 5 )
四川省卫生厅基金资助
关键词
免疫毫微粒
单克隆抗体
米托蒽醌
肝肿瘤
Nanoparticles Monoclonal antibody Mitoxantrone Human hepatoma
