摘要
目的制备分别携带VEGF和CD34抗体以及同时携带2种抗体的靶向超声造影剂,并对其体外寻靶能力及免疫活性进行鉴定。方法采用吸附法制造3种靶向超声造影剂,每种靶向超声造影剂又根据微泡与抗体的配比比例及pH值具体各分为6组,共18组(实验组)。使用荧光免疫法,在体外鉴定每个实验组的寻靶能力及免疫活性。同时,设普通造影剂对照组、二抗代替一抗对照组。在电镜下计算靶向超声造影剂的结合率,并进行比较。结果电镜及光镜下观察,靶向超声造影剂为圆形,大小及分布较均匀,无任何聚集现象。荧光显微镜下观察,MBV实验组、MBC实验组、MBB实验组与靶向超声微泡结合的内皮细胞胞浆表达为绿色,而未与靶向超声微泡结合的内皮细胞及其他实验组、普通造影剂对照组与二抗代替一抗对照组均未表达。电镜下观察,靶向超声微泡组部分内皮细胞与靶向微泡结合,MBV实验组的结合率为3.475 5%±0.139 2%,MBC实验组的结合率为3.544 4%±0.214 5%,MBB实验组的结合率为3.682%±0.124 6%,各组比较差异无统计学意义(P>0.05),而其他实验组及对照组未见与微泡结合的内皮细胞。结论自制的3种靶向超声造影剂在体外有寻靶能力及免疫活性,可用于动物肿瘤模型新生血管的体内特异性靶向显影的实验研究。
Objective To prepare targeted ultrasound contrast agents with the VEGF antibody,or the CD34 antibody,or both attached to microbubbles,and assess their targeting efficiency and immunoactivities.Methods 3 types of targeted ultrasound contrast agents were prepared using the absorption method.Each type of targeted ultrasound contrast agents were divided into 6 subgroups（18 in total）regarding the ratio of microbubbles to antibodies and pH values.The targeting efficiency and the immunoactivities were assessed in each group using the immunofluorescence method.Normal ultrasound contrast agents and the secondary antibodies replacing the primary antibodies were considered as the controls.The combination rate of targeted ultrasound contrast agents was calculated and compared under the electron microscope.Results Targeted ultrasound contrast agents were round,medium-sized,and they were distributed evenly without concentrations under the electron microscope and the light microscope.Under the fluorescence microscope,the cytoplasm of endothelial cells absorbing microbubbles of targeted ultrasound contrast agents was green in the MBV,MBC,and MBB groups,while there was no staining in the cytoplasm of endothelial cells in the endothelial cells not absorbing microbubbles,other treatment groups and groups with normal ultrasound contrast agents or the secondary antibodies replacing the primary antibodies.Under the electron microscope,the microbubbles of targeted ultrasound contrast agents were closely adherent to endothelial cells.The combination rates in the MBV,MBC,and MBB groups were 3.475 5%±0.139 2%,3.544 4%±0.214 5% and 3.682%±0.124 6% respectively.No significant difference was noted among them,P0.05.No endothelial cell adhered to microbubbles in other treatment groups and control groups.Conclusion The three types of targeted ultrasound contrast agents have targeting efficiency and immunoactivities in vitro and may be applied for specifically targeted imaging for neovessels in the animal tumor model.
出处
《实用药物与临床》
CAS
2011年第6期457-459,F0003,共4页
Practical Pharmacy and Clinical Remedies
