携Herceptin靶向高分子造影剂的制备及体外寻靶能力研究

目的 制备一种携Herceptin的靶向高分子造影剂,并研究其体外寻靶能力.方法 通过双乳化法制备出高分子PLGA-COOH造影剂,采用碳二亚胺法将高分子PLGA-COOH造影剂与Herceptin相连制备出靶向高分子造影剂,检测其一般性质,然后通过免疫荧光法检测其与抗体的连接情况,用光镜及激光共聚焦显微镜观察其与细胞的结合能力,并与普通高分子造影剂做比较.结果 靶向高分子造影剂的粒径范围为466.8～857.6 nm,平均粒径为(662.2±69.7)nm,有85.9%的微球粒径落于该范围内,通过免疫荧光法可在荧光显微镜下观察到许多绿色点状荧光,体外寻靶能力实验显示靶向高分子造影剂能与乳腺癌细胞较好的结合.结论 成功制备出的携Herceptin的靶向高分子造影剂在体外与人乳腺癌细胞有较好的结合能力.

Preparation and targeting study of Herceptin loaded PLGA-COOH-targeted ultrasound contrast agent

Objective To preparation a kind of Herceptin loaded and breast cancer targeted high molecular polymer ultrasound contrast agent and investigate their affinity for breast cancer in vitro. Methods The high molecular polymer PLGA-COOH ultrasound contrast agents were produced by the double emulsion technique. Herceptin was covalently linked to the PLGA-COOH nanoparticle surface using a carbodiimide technique. Its physical property was determined. The combination of Herceptin with the PLGA-COOH nanoparticle was proved by immunofluorescent assay,and the PLGA-COOH nanoparticle targeting specifity to breast cancer cells was observed with light microscope and confocal microscope,normal PLGA-COOH nanoparticle was served as control group. Results The diameter range of targeted high molecular polymer ultrasound contrast agents was 466.8～857.6 nm, the average diameter was (662.2 ± 69. 7) nm,85.9% of the diameter was in the range. Green punctiform fluorescence was observed by fluorescence microscope,and the conjugation of the targeted high molecular polymer ultrasound contrast agents with MCF-7 cells was tight while the conjugation in control group was negative. Conclusions The Herceptin loaded PLGA-COOH targeted ultrasound contrast agents was prepared successfully. The targeted ultrasound contrast agents can bind to breast cancer effectively in vitro.