亲和素桥连构建携抗P-选择素单抗靶向超声微泡及体外荧光鉴定  

Construction of targeted ultrasound microbubbles with anti P-selectin monoclone antibody via streptavidin bridge and evaluation of its reliability using fluorescence in vitro

作　　者：郑道文[1] 杨莉[2] 宾建平[1] 陈楚弟[3] 陈少敏[1] 王月刚[1] 吴平生[1] 

机构地区：[1]南方医科大学南方医院心内科,广东广州510515 [2]南方医科大学南方医院药学部,广东广州510515 [3]南方医科大学南方医院消化科,广东广州510515

出　　处：《中国医学影像技术》2008年第8期1182-1185,共4页Chinese Journal of Medical Imaging Technology

基　　金：国家"863"计划项目(2006AA02Z478)

摘　　要：目的体外荧光方法鉴定生物素-亲和素-生物素(BSB)桥连技术构建携带抗P-选择素单抗靶向超声微泡(MB-BSBp)的可靠性。方法不同荧光标记物分别标记MB-BSBp的不同部位,观测微泡荧光强度(0、1、2和3级),以普通脂质超声微泡(MB)作对照。结果加入FITC荧光亲和素孵育,生物素化脂质微泡(MB-B)呈现明亮的绿色荧光(3级),而MB无荧光显示(0级)。以两个浓度梯度(1∶4和1∶16)的DTAF荧光二抗(抗抗P-选择素单抗抗体)标记MB、MB-B、MB-BS和MB-BSBp,MB-BSBp在两个浓度梯度下均发出明亮的绿色荧光(3级);而MB-B、MB-BS仅在1∶4时显示微弱的绿色荧光(1级),MB在两个浓度梯度下均无荧光显示(0级)。结论抗P-选择素单抗通过亲和素桥连有效装配在MB-B表面,体外荧光法是鉴定靶向微泡配体连接可靠性的简便方法。Objective To investigate the reliability of targeted ultrasound microbubbles with anti-mouse P-selectin monoclone antibody（MB-BSBp）.Methods The targeted microbubbles were constructed using a technology of biotin-avidin-biotin（BSB）antibody conjugation.Different parts of MB-BSBp were marked with various fluorescent markers,and common lipid ultrasound microbubbles（MB）as control.The fluorescence intensity of microbubbles was graded from 0,1,2 to 3.Results After incubating with FITC-avidin,the biotinylated lipid microbubbles（MB-B）displayed bright green fluorescence（grade 3）,but MB had no any fluorescence（grade 0）.MB,MB-B,streptavidin-biotinylated lipid microbubbles（MB-BS）,and MB-BSBp were marked with DTAF antibody as second antibody（antibody of anti-P-selectin monoclonal antibody）at two concentrations（1∶4 and 1∶16 dilution）.Bright green fluorescence（grade 3）were observed in MB-BSBp in both concentrations,faint green fluoresence（grade 1）in MB-B and MB-BS only in concentration of 1∶4 dilution,but no fluorescence was seen in MB in either concentration（grade 0）.Conclusion P-selectin monoclonal antibody was effectively incorporated to the MB-B surface through streptavidin bridge.Fluorometric in vitro is reliable and simple method for investigating the ligand conjugation of targeted microbubble.

关 键 词：P-选择素 靶向超声微泡 荧光 

分 类 号：R445.1[医药卫生—影像医学与核医学]