CAJ | 학술논문

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-BAB-KDR，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。
목적탐토휴혈관내피생장인자수체2（KDR）단항적파향지질미포적제비방법，병대기물리특성、생물활성、파향점부은정성진행체외측정。방법채용성진의제비생물소화지질미포（MB-B），재응용생물소-친화소교련기술구건휴KDR단항적파향미포（MB-BAB-KDR）。형광현미경하관찰MB-B、단순단항미포（MB-B-KDR）、MB-BAB-KDR여형광이항부육후형광강도。이용평행판류동강기술체외모의생리혈류전절력조건，평개파향미포적점부효능。결과제비적파향미포MB-BAB-KDR정원구형，립도분포균균。여형광이항부육후，MB-BAB-KDR발출명량적록색형광（Ⅱ급），이MB-B-KDR현시미약적록색형광（Ⅰ급）、MB-B무형광（0급）。체외점부실험현시，MB-BAB-KDR결합수목수착소서KDR Fc포피농도적증고이증가（P<0.05），상동농도하，수착시간적증가，MB-BAB-KDR결합수목증가。결론성공구건생물소-친화소교련적파향지질체미포MB-BAB-KDR，체외점부실험현시MB-BAB-KDR구유점부은정성，차여시간、농도정정상관。
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.