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目的研究人鼠嵌合抗肿瘤细胞核单抗(chTNT-3)-空间稳定脂质体的制备方法、免疫活性及体外靶向性。方法合成聚乙二醇末端带吡啶二硫丙酰基的磷脂衍生物(PDP-PEG-HSPE),制备含PDP-PEG-HSPE的空间稳定脂质体,经二巯基苏糖醇还原后共价连接马来酰亚胺衍生化抗体。荧光胺法和钼蓝法测定脂质体与抗体的连接效率及抗体密度,激光散射粒度仪测定其粒径分布,ELISA法检测脂质体表面的抗体免疫活性。体外实验考察该免疫脂质体与固定Raji细胞的结合活性。结果chTNT-3-空间稳定脂质体的粒径分布为(115±33)nm。当初始Ab/PDP-PEG-HSPE=1∶10时,脂质体与抗体的连接效率为71%,抗体密度为106μgAb/μmolPL。chTNT-3经化学修饰后连接到脂质体表面,其免疫活性基本保留。chTNT-3-空间稳定脂质体能特异性地结合固定Raji细胞。结论通过PDP-PEG-HSPE法共价连接抗体制备的chTNT-3-空间稳定脂质体能基本保留chTNT-3的免疫活性,具有体外靶向细胞核抗原的能力。
Objective To study the preparation, immunological activity and in vitro targeting of human chimeric antitumor cell nuclear monoclonal antibody (chTNT-3) -specific stable liposome. Methods The PEG-PEG-HSPE derivatives were synthesized at the terminal of polyethylene glycol (PEG-PEG-HSPE), and the stable liposomes containing PDP-PEG-HSPE were prepared and reduced by dimercaptothreitol Maleimide Derivatized Antibodies. Fluorescent amine method and molybdenum blue method were used to determine the liposome-antibody ligation efficiency and antibody density. The particle size distribution was measured by laser scattering particle size analyzer. The immunological activity of the liposome was detected by ELISA. The in vitro experiments investigated the binding activity of this immunoliposome to immobilized Raji cells. Results The size distribution of chTNT-3-stable steroid liposomes was (115 ± 33) nm. When the initial Ab / PDP-PEG-HSPE = 1: 10, the liposome-to-antibody ligation efficiency was 71% and the antibody density was 106 μgAb / μmol PL. ChTNT-3 chemically modified to connect to the surface of the liposome, its immunological activity remained essentially. ChTNT-3-sterically stabilized liposomes specifically bind to immobilized Raji cells. Conclusion The chTNT-3-sterically stabilized liposomes prepared by the covalent attachment of antibodies by the PDP-PEG-HSPE method can substantially retain the immunogenicity of chTNT-3 and have the ability to target nuclear antigen in vitro.