静电吸附法与生物素-亲和素桥联法制备PSMA抗体靶向纳米微泡的对比实验研究  

Contrast experimental study on preparation of PSMA-targeted nanobubbles using electrostatic adsorption method and biotin avidin-bridging technology

作　　者：李浪 郭燕丽 张淼 范校周 童海鹏 

机构地区：[1]第三军医大学第一附属医院超声科,重庆市400038

出　　处：《临床超声医学杂志》2013年第12期809-812,共4页Journal of Clinical Ultrasound in Medicine

基　　金：国家自然科学基金(30970830);重庆市科委自然科学基金(cstc2012jjB10072)

摘　　要：目的比较静电吸附法和生物素-亲和素桥联法制备前列腺特异性膜抗原(PSMA)抗体靶向纳米微泡的物理特性和靶向黏附率。方法分别应用静电吸附法和生物素-亲和素桥联法制备PSMA抗体靶向的纳米微泡,检测其物理性状;光镜下观察两种微泡与前列腺癌细胞LNCaP和C4-2的靶向能力,并与胃癌细胞MKN45对照;对比分析两种靶向纳米微泡的粒径、浓度及与前列腺癌细胞结合能力。结果两种靶向纳米微泡均呈圆形,表面光滑,大小均匀,分散度好,无聚集,粒径均<1000 nm。静电吸附法制备的靶向纳米微泡平均粒径为(702.96±66.65)nm,平均浓度为(3.46±0.30)×108/ml,与前列腺癌细胞LNCaP黏附率(77.2±3.19)%,与C4-2细胞黏附率(61.00±4.47)%;生物素-亲和素桥联法制备的靶向纳米微泡平均粒径为(609.90±37.40)nm,平均浓度约(4.52±0.19)×108/ml,与前列腺癌细胞LNCaP黏附率(96.60±1.14)%,与C4-2细胞黏附率(94.00±1.58)%。两种靶向纳米微泡的粒径、浓度及与前列腺癌细胞黏附率比较,差异均有统计学意义(均P<0.05);与胃癌细胞MKN45均无黏附。结论生物素-亲和素桥联法是一种高效、灵敏、稳定、特异性强的PSMA抗体靶向微泡制备方法,较静电吸附法具有更大的优势。Objective To compare the characteristics and adhesion rate of PSMA-targeted nanobubbles prepared using electrostatic adsorption method and biotin avidin-bridging technology. Methods PSMA monoclonal antibody-loaded targeted nanobubbles were prepared using electrostatic adsorption technology and biotin-avidin bridging technology. Their general characteristics were detected. Using MKN45 gastric cancer cells as the control, the targeting capability of the targeted nanobubbles to prostate cancer cells （LNCaP and C4-2） was observed under optical microscope. The differences between the two kinds of targeted nanobubbles in particle size, stability, and the ability adhered to prostate cancer cells was analyzed. Results The two kinds of targeted nanobubbles were round and had the uniform size with good dispersity and no aggregation. The average particle size of PSMA-targeted nanobubble prepared with electrostatic adsorption was （702.96±66.65）nm, concentration was about （3.46_＋0.30） ×10^8/ml, and prostate cancer cells LNCaP adhesion rate was （77.2±3.19）%, and C4-2 cell adhesion rate was （61.00±4.47 ）%; while the average particle size of PSMA-targeted nanobubble prepared with avidinbiotin complex technology was （609.90±37.40）nm, concentration was about （4.52±0.19） ×10^8/ml, and prostate cancer cells LNCaP adhesion rate was （ 96.60± 1.14）%, and C4-2 cell adhesion rate was （94.00±1.58）%. The differences of particle size, concentration and cell adhesion rate of the two kinds of targeted microbubble were statistically significant（P〈0.05 ）. The two kinds of nanobubbles were not adhered to MKN45 gastric cancer cells. Conclusion Biotin-avidin bridging method has more advantages than electrostatic adsorption method in preparation of PSMA-targeted nanobubbles. It is an efficient, sensitive, stable, specific method.

关 键 词：靶向微泡 前列腺特异性膜抗原 静电吸附法 生物素-亲和素系统 

分 类 号：R737.25[医药卫生—肿瘤]