携IgG单抗靶向超声造影剂的构建及其靶向效能的体外评价  

Construction of targeted ultrasound contraSt agent with anti-IgG monoclone antibody and the evaluation on the effectiveness of its targeted adhesion in vitro

作　　者：李军华[1] 吴爵非[1] 杨莉[2] 刘俭[1] 郑道文[1] 纪丽景[1] 燕翼[1] 宾建平[1] 

机构地区：[1]南方医科大学南方医院心内科,广州510515 [2]南方医科大学南方医院药学部,广州510515

出　　处：《中华超声影像学杂志》2009年第9期813-816,共4页Chinese Journal of Ultrasonography

基　　金：国家863计划项目（2006AA022478）

摘　　要：目的构建携羊抗小鼠IgG单抗靶向超声造影剂（UCA-IgG）及体外评价其靶向黏附效能。方法采用“亲和素一生物素”桥接法构建UCAIgG，体外荧光法鉴定IgG单抗与生物素化造影剂特异结合。利用平行板流动腔在不同时间点、不同浓度小鼠IgG包被和不同剪切应力下，测定相应的UCA-IgG的结合数目及达到半数解离时的剪切应力。设不同浓度小鼠包被的平行板流动腔为实验组，羊抗小鼠IgG封闭组和空白组为对照组。结果UCAIgG发出明亮的绿色荧光，而普通脂质造影剂无荧光显示。实验组UCA-IgG结合数量随包被浓度的增高而增加（P〈0．05），并与结合时间呈明显正相关（P〈0．05），然而与剪切应力呈现双向性（P〈0．05）。而两对照组均未见明显的UCAIgG结合。UCA-IgG达半数解离的剪切应力随包被浓度的增加而增大（P〈0．05）。结论在不同血流动力学条件下UCA-IgG可与小鼠IgG特异有效结合，携羊抗小鼠IgG单抗造影剂可为其他特异靶向超声造影剂的体外研究提供有力的支持。Objective To construct targeted ultrasound contrast agent carried goat anti mouse IgG antibody （UCA-IgG） and evaluate the effectiveness of its targeted adhesion using parallel plate flow chamber. Methods The ultrasound contrast agent targeted to mouse IgG was designed by conjugating monoclonal antibodies against mouse IgG to the lipid monolayer shell of the agent using biotin-streptavidin. The binding of IgG antibodies to the ultrasound contrast agent were identified by fluorescence in vitro. The attachment and detachment of UCA-IgG to mouse IgG immobilized on a culture dish were assessed in a parallel plate flow chamber. While the plate lacked mouse IgG,or blocked with large number of goat anti mouse IgG were served as two control groups. Results UCA IgG issued a bright green fluorescence, while the contral lipid ultrasound contrast agent didn＇t show fluorescence. The number of UCA-IgG bound to mouse IgG of experimental group was greater than two control groups, increased with increasing coverslips surface antibody concentrations （ P 〈0.05）, and there was significant positive correlation between the number of UCA-IgG bound to mouse IgG and time of combination （ P 〈0.05）. The adhesion rate of experimental group increased with shear stress before 0.5 ×10 ^-5N/cm^2 （ p 〈 0.05） and then decreased （ P 〈0.05）. There was limited adherence of control groups to the UCA-IgG. The stess of half maximal detachment was increased with increasing eoverslips surface antibody concentrations （ P 〈0.05）. Conclusions UCA-IgG could adhere to mouse IgG in the physical conditions. It may provide strong supports for studying other targeted ultrasound contrast agent preliminary and fatherly in vitro.

关 键 词：超声检查 微气泡 靶基因修复 免疫球蛋白G 

分 类 号：R686[医药卫生—骨科学]