上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

携抗HER2抗体和抗VEGFR2抗体聚乳酸羟基乙酸双靶向纳米高分子超声造影剂制备及体外实验研究

李晓钰1，杜晶1，杨仕平2，胡鹤2，李凤华1   

  1. 1.上海交通大学 医学院附属仁济医院超声医学科， 上海 200127； 2.上海师范大学 化学系， 上海 200234
  • 出版日期:2016-01-28 发布日期:2016-02-26
  • 通讯作者: 李凤华, 电子信箱: fenghua-li@163.com。
  • 作者简介:李晓钰（1988—）, 女， 硕士生; 电子信箱: xiaoyuli37@163.com。
  • 基金资助:

    国家自然科学基金（81102014）

Preparation and in vitro experimental study on poly lactic-co-glycolic acid dual-targeted nanoparticle polymer ultrasound contrast agent carrying both anti-HER2 and anti-VEGFR2 antibodies

LI Xiao-yu1, DU Jing1, YANG Shi-ping2, HU He2, LI Feng-hua1   

  1. 1.Department of Ultrasound， Renji Hospital， Shanghai Jiao Tong University School of Medicine， Shanghai 200127， China; 2.Department of Chemistry， Shanghai Normal University， Shanghai 200234， China
  • Online:2016-01-28 Published:2016-02-26
  • Supported by:

    National Natural Science Foundation of China, 81102014。

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摘要：

目的 制备携抗人上皮生长因子受体2(HER2)抗体和抗血管内皮细胞生长因子受体2（VEGFR2）抗体的聚乳酸羟基乙酸(PLGA)双靶向超声造影剂，观察其在体外寻靶的能力和超声显像效果。方法 通过改进的双乳化溶剂挥发法制备PLGA纳米粒子，利用透射电子显微镜及激光粒度仪对其一般特性进行分析；利用碳二亚胺法将抗HER2抗体和抗VEGFR2抗体与PLGA粒子共价结合，制备出同时携带抗HER2抗体和抗VEGFR2抗体的双靶向纳米高分子超声造影剂。用激光共聚焦扫描显微镜评估其体外寻靶能力，流式细胞仪检测其与靶向细胞结合率，并使用高频超声诊断仪观察其体外显像效果。结果 PLGA高分子纳米超声造影粒子平均粒径为（152.00±58.08）nm，呈球形,表面较光滑,分散性好,具有空心结构。体外寻靶实验显示，携抗HER2抗体和抗VEGFR2抗体的PLGA双靶向造影剂较多并牢固地聚集到乳腺癌细胞SKBR3和血管肉瘤内皮细胞SVR表面。体外超声成像实验显示，PLGA双靶向纳米超声造影剂呈细腻均匀的点状密集高回声，后方未见衰减回声。结论 成功制备出同时携带抗HER2抗体和抗VEGFR2抗体的PLGA双靶向纳米高分子造影剂。该造影剂在体外能与高表达HER2受体的SKBR3细胞和高表达VEGFR2受体的SVR细胞特异性结合，并且具有良好的超声体外显影效果。

关键词: 聚乳酸羟基乙酸, 靶向超声造影剂, 人上皮生长因子受体2, 血管内皮生长因子受体2

Abstract:

Objective To prepare the poly(lactic-co-glycolic acid)(PLGA) dual-targeted ultrasound contrast agent carrying anti-human epidermal growth factor receptor 2 (anti-HER2) and anti-vascular endothelial growth factor receptor 2 (anti-VEGFR2) antibodies and observe the targeting ability and the effect of ultrasound imaging in vitro. Methods The PLGA nanoparticles (NPs) were prepared by a modified double emulsious-solvent evaporation technique and the general characteristics of PLGA NPs were analyzed by the transmission electron microscope and laser particle size analyzer. The anti-HER2 and anti-VEGFR2 antibodies were conjugated to PLGA NPs by the carbodiimide method to prepare the dual-targeted polymer nanoparticle ultrasound contrast agent carrying both anti-HER2 and anti-VEGFR2 antibodies. The targeting ability in vitro was assessed by the laser scanning confocal microscope (LSCM) and the rate of combination with target cells was detected by flow cytometer. The effect of imaging in vitro was observed by the high frequency ultrasonography. Results The PLGA NPs were round with smooth surface, good dispersion, and hollow structure. The average particle size was (152.00±58.08) nm. The results of targeting tests in vitro showed that the PLGA dual-targeted NPs carrying anti-HER2 and anti-VEGFR2 antibodies were firmly attached to the surface of human breast cancer cells SKBR3 and endothelial cells of angiosarcoma SVEN1 ras (SVR). The results of ultrasonic imaging tests in vitro indicated that the PLGA dual-targeted nanoparticle ultrasound contrast agent presented uniform and fine dotted dense high echo with no attenuated echo in the rear area.  Conclusion The PLGA dual-targeted nanoparticle polymer ultrasound contrast agent carrying both anti-HER2 and anti-VEGFR2 antibodies is successfully prepared. This contrast agent can specifically bind to SKBR3 cells with high expression of HER2 and SVR cells with high expression of VEGFR2 in vitro. The effect of ultrasound imaging in vitro is satisfactory.

Key words: poly(lactic-co-glycolic acid), targeted ultrasound contrast agent, human epidermal growth factor receptor 2, vascular endothelial growth factor receptor 2