抗人肝癌免疫毫微粒的制备及体外免疫学性质的鉴定

目的 构建人肝癌特异的免疫毫微粒,探讨免疫微粒体外对靶细胞的特异性结合特性及杀伤性活。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

Preparation and immunological characterization of mitoxantrone-loaded immuno-nanoparticles against human hepatoma

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.