CAJ | 학술논문

目的制备以HER2受体为靶点的靶向纳米级脂质微泡超声造影剂,并观察其体外寻靶能力及体外超声显像效果.方法制备生物素化Herceptin单抗,检测其生物素化程度及生物学活性；薄膜水化-声振法制备生物素化纳米微泡,以生物素-亲和素为桥梁制备HER2为靶点的靶向纳米级脂质微泡超声造影剂,观察其对SKOV3卵巢癌细胞的体外寻靶能力及靶向结合的体外超声显像.结果平均每分子Herceptin单抗可与16个生物素分子结合；与游离单抗相比,生物素化抗体的活性未见明显降低(P＞0.05).体外寻靶实验观察:靶向纳米微泡组SKOV3细胞表面有明显的红染纳米微泡与其牢固结合,沿细胞表面排列较规则；非靶向组SKOV3细胞表面未结合红染的纳米微泡.靶向纳米微泡体外超声显像:细胞爬片与靶向纳米微泡孵育后可明显增强超声显像效果,对照组均未见明显超声显像.结论应用生物素-亲和素系统成功构建了以HER2为靶点的纳米级超声造影剂,与SKOV3细胞结合后有明显超声显像效果.
목적 제비이HER2수체위파점적파향납미급지질미포초성조영제,병관찰기체외심파능력급체외초성현상효과.방법 제비생물소화Herceptin단항,검측기생물소화정도급생물학활성；박막수화-성진법제비생물소화납미미포,이생물소-친화소위교량제비HER2위파점적파향납미급지질미포초성조영제,관찰기대SKOV3란소암세포적체외심파능력급파향결합적체외초성현상.결과 평균매분자Herceptin단항가여16개생물소분자결합；여유리단항상비,생물소화항체적활성미견명현강저(P＞0.05).체외심파실험관찰:파향납미미포조SKOV3세포표면유명현적홍염납미미포여기뢰고결합,연세포표면배렬교규칙；비파향조SKOV3세포표면미결합홍염적납미미포.파향납미미포체외초성현상:세포파편여파향납미미포부육후가명현증강초성현상효과,대조조균미견명현초성현상.결론 응용생물소-친화소계통성공구건료이HER2위파점적납미급초성조영제,여SKOV3세포결합후유명현초성현상효과.
Objective To prepare targeted nanoscale lipid ultrasound contrast agent and study the targeting function in vitro.Methods After the biotinylated monoclonal antibody Herceptin was prepared,the biotinylated degree and immunological activity were determined.Then biotinylated antibody was attached to the surface of nanoscale lipid ultrasound contrast agents by avidin-biotin system to prepare the targeted nanobubbles.The targeting function was studied by observing the combination ability of the targeted nanobubbles with SKOV3 cells in vitro,non-targeted nanobubbles as controls,and observing ultrasound imaging in vitro.Results About 16 biotin molecules were coupled to each antibody in average,and the immunological activity of the biotinylated antibody didn't decrease compared with the free one(P ＞0.05).SKOV3 cells were combined firmly and surrounded regularly by red dyed targeted nanobubbles,while control groups were negative.Ultrasound imaging could be significantly enhanced by targeted nanobubble binding to SKOV3 cell slides,the other two control groups were negative.Conclusions Nanoscale ultrasound contrast agent and antibodys can be combined firmly by avidin-biotin system to produce the targeted nanobubbles,which have strong targeting function in vitro and significantly enhanced ultrasound signal.