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抗人肝癌免疫毫微粒的制备及体外免疫学性质的鉴定 |
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刘晓波 蔡美英
摘 要 目的：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SP DP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-7721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。
关键词 毫微粒 单克隆抗体 人肝癌 米托蒽醌
中国图书分类号 R392.2 R730.61
Preparation and immunological characterization of mitoxantrone-loaded immuno-nanoparticles against human hepatoma
LIU Xiao-Bo CAI Mei-Ying
（Department of Immunology, West China University of Medical Sciences, Chengdu 610041）
Abstract Objective：To construct human hepatoma-specific immunonanoparticles and observe in vitro binding and killing of target cells by the immunonanoparticles.Methods:Hetero bifunctional cro sslinker SPDP was used to couple covalently McAb HAb18(anti-human hepatoma) wit h mitoxantrone-loaded bovine serum albumin nanoparticles(DHAQ-BSA-NP). A lot o f in vitro experiments,such as slide agglutination test,immunofluorescent assay, i mmunofluorescent blocking test,rosset formation test,rosset formation blocking t est and scanning electron microscopy etc,were used to confirm coupling of antibo dies to nanoparticles and specific binding of immunonanoparticles with target c e lls.3 H-TdR incorporation test was used to assay cytotoxicity of immunonano par ticles to target cells.Results:McAb HAb18 was coupled to D HAQ-BSA-NP. The immunonanoparticles could bind specifically to human hepatoma SMMC-7721 cells and exert selective killing effects on the target cells with do se-dependence.Conclusion:Conjugation technique via SPDP i s a good method for constructing immunonanoparticles. Human hepatoma-specific- immunonano particles have capacity of in vitro selective coating and killing human hepatoma cell line.
Key words Nanoparticles Monoclonal antibody Mitoxantrone Human hepatoma
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