流式细胞术定量评价携抗P-选择素单抗靶向超声微泡配体结合效率  

Quantitative Evaluation on the Bonding Efficiency of the Ligands on the Microbubb｜es Targeted to P-selectin by Flow Cytometry

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

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

出　　处：《中国超声医学杂志》2009年第7期632-635,共4页Chinese Journal of Ultrasound in Medicine

基　　金：本课题受国家“863”计划项目（No.2006AA022478）资助

摘　　要：目的流式细胞术定量评价应用生物素（B）-亲和素（S）-生物素（B）桥连技术构建的携抗P-选择素靶向微泡（M辟BSBp）的配体结合效率。方法以未标记荧光的MB—B作对照，用不同荧光物质分别标记MB-BSBp不同部位，应用流式细胞仪分析各组微泡的荧光强度并定量配体结合率。C37小鼠8只建立肾脏缺血再灌注模型载体评价靶向微泡成像质量。结果MB-B与FITC—streptavidin的结合率达99％；DTAF-二抗与MB-BSBp的结合率为（59．66±2．30）％，明显高出荧光二抗与MB-BS和MB-B的结合，比率分别为（29．87±1．68）％和（26．50±3．86）％（P〈0．05）。结论抗P-选择素单抗通过生物素桥连法有效装配在MB-B表面，流式细胞术可能是定量评价靶向微泡配体结合效率的有效方法。Objective To investigate the bonding efficiency of the ligands on the microbubbles with anti-mouse P-selectin monoclone antibody （MB-BSBP） by flow cytometry quantitatively. Methods Different ligands of the MB-BSBP were marked by different fluorescence respectively. Banding efficiency of the ligands was quantified via the fluorescent intensity and assessed by flow cytometry. MB-B without marks was served as control. Renal isehemia-reperfusion （IR） model was created with 8 C37 mice and then the contrast-enhanced ultrasound （CEU） imaging by MB-BSBp was performed. Results 99% of MB-B was incorporated with FITc streptavidin. The bonding efficiency of DTAF to MB-BSBp was （59.66 ± 2.30）%, and it was much higher than that to MB-BS and MB-B, which were （29.87± 1.68）% and （26.50±3.86）% respectively （P〈0.05） . Conclusions P selectin monoelonal antibody was effectively linked to the MB-B surface through streptavidin bridge. Flow cytometry could be a reliable quantitative method to investigate the bonding efficiency of the ligands on the microbubbles.

关 键 词：靶向超声微泡 流式细胞术 对比超声 P-选择素 

分 类 号：R692.3[医药卫生—泌尿科学] R733.71[医药卫生—外科学]