肝癌靶向液态氟碳脂质微球造影剂的体外寻靶实验研究

Targeting study in vitro of human hepatocellular carcinoma-targeted perfluorocarbon lipid particles

目的制备液态氟碳脂质微球造影剂并研究其体外基本特性，再引入抗人肝癌细胞单克隆抗体，采用生物素亲和素法，进行体外靶向结合的研究。方法采用旋转蒸发法和高压均质法制备一种纳米液态氟碳脂质微球造影剂，观察其外观、形态、大小及分布情况，计算其浓度，并测定其粒径、电位；制备生物素的抗人肝癌细胞单克隆抗体HAb18，用HABA／Avidin试剂盒检测抗体的生物素化程度；制备NBD标记的生物素化的微球，采用生物素亲和素法研究其体外靶向作用。结果所制备的液态氟碳脂质微球造影剂外观呈乳白色的混悬液，镜下观察，微球形态圆整，大小均一，性质稳定，平均粒径171．9nm；采用生物素亲和素法制备的NBD标记的生物素化的微球可靶向结合到靶细胞上。结论成功制备出稳定的纳米液态氟碳脂质微球造影剂，并引入抗人肝癌细胞单克隆抗体，通过生物素亲和素系统可将其特异结合到靶细胞上。

Objective To develop perfluorocarbon lipid particles and investigate their basic properties,and target them to human hepatocellular carcinoma cells in vitro by hepatoma monocolonal antibody HAb18 with avidin-biotin interaction. Methods Rotary evaporation and high pressure homogen were used to prepare perfluorocarbon lipid particles, and the appearance and distribution of them were investigated by microscope and electron microscope, the concentration and the size and electric potential were detected. The biotinylated monoclonal antibody HAb18 was prepared, then the biotinylated degree of the antibody was determined. The biotinylated perfluorocarbon lipid particles labelled with NBD were prepared and targeted to human hepatocellular carcinoma cells in vitro with avidin-biotin interaction. Results These perfluorocarbon lipid nanoparticles were uniform and stable,and the mean diameter of them was 171.9 nm. Hepatocellular carcinoma cells were surrounded by the biotinylated particles labelled with NBD. Conclusions A steady perfluorocarbon lipid particles were prepared and the biotinylated particles can be targeted to hepatocellular carcinoma cells with avidin-biotin interaction.