血管内皮生长因子受体2单抗的靶向超声造影剂制备及体外评价  

Detection of binding capability of targeted KDR ultrasound contrast agent in vitro for evaluating endometrial receptivity

作　　者：刘红梅[1] 韩小华[1] 杨莉[2] 纪丽景[3] 李素淑[1] 

机构地区：[1]南方医科大学第三附属医院超声医学科,广东广州510630 [2]南方医科大学南方医院药剂科,广东广州510515 [3]南方医科大学南方医院超声科,广东广州510515

出　　处：《南方医科大学学报》2013年第9期1308-1311,共4页Journal of Southern Medical University

基　　金：国家自然科学基金(81101063);广东省自然科学基金(S2012010008115);广东省科技计划项目(2010B031600244)~~

摘　　要：目的探讨携血管内皮生长因子受体2(KDR)单抗的靶向脂质微泡的制备方法,并对其物理特性、生物活性、靶向黏附稳定性进行体外测定。方法采用声振仪制备生物素化脂质微泡(MB-B),再应用生物素-亲和素桥连技术构建携KDR单抗的靶向微泡(MB-BAB-KDR)。荧光显微镜下观察MB-B、单纯单抗微泡(MB-B-KDR)、MB-BAB-KDR与荧光二抗孵育后荧光强度。利用平行板流动腔技术体外模拟生理血流剪切力条件,评价靶向微泡的黏附效能。结果制备的靶向微泡MB-BAB-KDR呈圆球形,粒度分布均匀。与荧光二抗孵育后,MB-BAB-KDR发出明亮的绿色荧光(Ⅱ级),而MB-B-KDR显示微弱的绿色荧光(Ⅰ级)、MB-B无荧光(0级)。体外黏附实验显示,MB-BAB-KDR结合数目随着小鼠KDR Fc包被浓度的增高而增加(P<0.05),相同浓度下,随着时间的增加,MB-BAB-KDR结合数目增加。结论成功构建生物素-亲和素桥连的靶向脂质体微泡MB-BABKDR,体外黏附实验显示MB-BAB-KDR具有黏附稳定性,且与时间、浓度呈正相关。Objective To prepare a new targeted liposome ultrasonic contrast agent with anti-KDR antibody that binds specifically with KDR as the main receptor of VEGF and evaluate its physical characteristics, biological activity and specific binding capability in vitro. Methods A sonicator was used to prepare the biotinylated lipid micro-bubbles （MB-B）, and biotin-avidin-mediated technique was used for attachment of anti-mouse KDR monoclonal antibody to the micro-bubble shell to generate MB-BAB-KDR. MB-BAB-KDR was incubated with fluorescent second antibody to assess the link condition, and the control groups were the MB-B and micro-bubbles with the antibody alone （MB-B-KDR）. A parallel plate flow chamber system was used to characterize micro-bubbles attachment efficiency to KDR Fc. Results The surface of the micro-bubbles could carry KDR antibody through the biotin-avidin bridge and MB-BAB-KDR were spherical and well-distributed. After incubation with the second antibody, MB-BAB-KDR could be observed to emit bright green fluorescence （Grade II） as compared with little or weak fluorescence in the control MB-B group （Grade 0） and MB-B-KDR group （Grade I）. Targeted micro-bubbles bound to KDR Fc increased as the KDR Fc concentration increased （P〈0.05）. Conclusion The targeted liposome contrast agent linked with KDR antibody by biotin-avidin bridge we prepared shows an increased binding number as the KDR Fc concentration increases, which provides a novel approach to molecular imaging study of endometrial receptivity.

关 键 词：靶向超声造影剂 血管内皮生长因子受体2 平行板流动腔 子宫内膜容受性 黏附 

分 类 号：R714.8[医药卫生—妇产科学]