载ICAM-1抗体的靶向脂质纳米超声造影剂体外寻靶实验研究  

In Vitro Evaluation of Targeting Capability of Targeted Lipid Nanoparticles Combining Anti-ICAM-1 Monoclone Antibody

作　　者：李文艳[1] 王志刚[1] 汪朝霞[1] 刘兴钊[1] 

机构地区：[1]重庆医科大学超声影像研究所,重庆市400010

出　　处：《中国超声医学杂志》2013年第2期173-176,共4页Chinese Journal of Ultrasound in Medicine

基　　金：国家自然科学基金青年科学基金项目(No.81101054);国家自然科学基金面上项目(No.81071126);国家自然科学基金重点项目(No.81130025);重庆高校优秀成果转化项目(No.KJZH10201)

摘　　要：目的制备一种载细胞间黏附因子-1(ICAM-1)单克隆抗体的脂质纳米超声造影剂,研究其体外寻靶能力。方法采用机械振荡法制得普通脂质纳米超声造影剂和生物素化的脂质纳米超声造影剂,利用生物素亲和素桥接法将ICAM-1抗体连接于脂质纳米超声造影剂上,制成靶向造影剂,检测两种造影剂的物理性质。将上述两种造影剂分别作用于普通HepG2细胞和经TNF-α处理的HepG2细胞,实时荧光定量PCR检测HepG2细胞上ICAM-1荧光表达强度,荧光显微镜观察造影剂与HepG2细胞靶向结合的情况。结果普通造影剂粒径为(623.2±91.37)nm;靶向造影剂粒径为(760.6±145.0)nm。经TNF-α刺激后HepG2细胞ICAM-1基因的表达量明显增高。普通造影剂组均未见造影剂与HepG2结合,而靶向造影剂组可见大量的造影剂聚集在经TNFα刺激后的HepG2细胞周围。结论本研究自制的载ICAM-1抗体的靶向脂质纳米超声造影剂,其粒径小,性质稳定,在体外能与高表达ICAM-1的HepG2细胞特异结合,可望成为一种良好的肝癌靶向造影剂。Objective To construct targeted lipid nanoparticles combining the anti-human ICAM-1 moloelone antibodies to the shell via＂avidin-biotin＂bridging chemistry and to evaluate its ability of targeting to HepG2 ceils. Methods Common lipid nanoparticles and biotined lipid nanoparticles were prepared by perfluorocarbon gas with several lipids in a certain ratio. Then more avidin was added to incorporate to the biotined nanoparticles. After being washed to remove excess avidin, a dose of biotin mouse-anti human ICAM-1 monoclone antibodies was added to complete the preparation of targeted lipid nanoparticles. Detect the partical size and electric potential Add the two contrast agents to groups HepG2 cells, wherein a group of cells after TNF a treatment. Real-time PCR detected ICAM 1 fluorescence strength of HepG2 cells and the ability of lipid nanoparticles targeting to HepG2 was observed on fluorescent microscopy. Results Common nanoparticles partical size were（623.2±91.37）nm, targeted nanoparticles partical size were （760.6±145.0）nm. Both of C＋P and C＋P-Ab groups had little green nanoparticles adhere to HepG2. A great quan- tity of green nanoparticles targeted to HepG2 in S＋P-Ab group ,which was higher than S＋P group dramatically. Conclusions This study constructed a type of targeted lipid nanoparticles conjugating mouse-anti human ICAM 1 mon oclone antibodies on the shell successfully, which has little partical size and was appropriate for targeting HepG2 in vitro.

关 键 词：脂质纳米超声造影剂 靶向超声分子成像 细胞间黏附因子一1 

分 类 号：R445.1[医药卫生—影像医学与核医学]