携抗ICAM-1单抗和抗CD34单抗双靶向微泡的制备及体外寻靶研究  

Dual-targeted microbubbles carrying both anti-CD34 monoclonal antibody and anti-ICAM-1 monoclonal antibody： preparation and investigation of its targeting performance in vitro

作　　者：叶露薇[1] 王志刚[1] 张萍[1] 岳媛媛[1] 

机构地区：[1]重庆医科大学附属第二医院超声影像学研究所,400010

出　　处：《中华超声影像学杂志》2013年第7期615-620,共6页Chinese Journal of Ultrasonography

基　　金：重庆市教委科学技术研究项目（KJ110320）

摘　　要：目的制备同时携抗ICAM-1单抗和抗CD34单抗的双靶向微泡，鉴定其基本性质，并观察体外寻靶能力。方法采用“生物素-亲和素”桥连法分别构建携抗ICAM-1单抗和抗CD34单抗双配体（MBD）、携抗ICAM-1单抗（MBICAM-1）和携抗CD34单抗（MBCD34）3种靶向微泡，光学显微镜下观察靶向微泡的形状，并用马尔文激光粒度分析仪、DFY超声图像定量分析仪、激光共聚焦显微镜、流式细胞仪检测靶向微泡的基本特性和抗体结合率。倒置显微镜下观察双靶向微泡与内皮祖细胞（EPCs）和损伤人脐静脉内皮细胞（HUVECs）的结合情况。结果3种靶向微泡的形状、粒径、表面电位及抗体结合率差异均无统计学意义（P〉0．05），MBD的回声强度高于MBICAM-1、MBCD34和普通生物素化微泡（MBBiotin）（P〈0．01）。MBD与EPCs和损伤HUVECs的结合率明显高于MBBiotin（P〈0．01），MBD和MBCD34与EPCs的结合率、MBo和MBmm与损伤HUVECs的结合率差异均无统计学意义（P〉0．05）。结论成功制备了携抗ICAM-1单抗和抗CD34单抗的双靶向微泡，体外实验证实此双靶向微泡与内皮祖细胞和损伤血管内皮细胞均能特异性结合。Objective To prepare dual-targeted microbubbles carrying both anti-CD34 monoclonal antibody and anti-ICAM-1 monoclonal antibody, to identify its basic characteristics, and to investigate its targeting performance in vitro. Methods ICAM-l-targeted microbubbles （MBICAM-1）, CD34-targeted microbubbles（MBCD34 ） and dual-targeted （with both monoclonal antibodies） microbubbles （MBo） were prepared by biotin-avidin bridging chemistry method. The targeted microbubbles were observed under light microscrope and characterized by a Mastersizer 3000, DFY software, laser confocal microscopy, and flow cytometry. The targeting specificity and attachment capability of dual-targeted microbubbles to endothelial progenitor cells （EPCs） and damaged human umbilical vein endothelial cells （HUVECs） were assessed in vitro. Results The morphous, diameter, surface potential, concentration, and the antibody-binding rates of microbubbles were not statistically significant in MBICAM-1 , MBCD34, and MBD （ P〉0.05）. Echo intensity in MBD significantly increased compared with MBICAM-1 , MBCD3,, and MBBiotin（ P〈 0.01 ）. The targeting experiment in vitro showed that the attachment rate of MBD to EPCs and damaged HUVECs significantly increased compared with MBBiotin（ P〈0.01）, while no remarkable difference in attachment rate was found between MBD and MBcD34 in EPCs, and between MBD and MBICAM-1 in damaged HUVECs （ P〉0.05）. Conclusions Dual-targeted microbubbles carrying both anti-CD34 monoclonal antibody and anti-ICAM-1 monoclonal antibody are prepared successfully. The study in vitro has proved that the dual-targeted microbubbles can specifically bind to both EPCs and damaged HUVECs.

关 键 词：超声检查 微气泡 内皮细胞 胞间黏附分子1 

分 类 号：R445.1[医药卫生—影像医学与核医学]