肿瘤新生淋巴管靶向脂质微泡的制备及超声显像研究

Targetting microbubbles of tumor lymphangiogenesis and ultrasound contrast imaging in vivo

目的：优化制备靶向到肿瘤新生淋巴细胞的抗体高携载率脂质超声微泡,即载 VEGFR-3抗体和 Podoplanin 抗体的高效靶向微泡,观察荷瘤模型超声显影效果。方法生物素-亲和素桥接法制备载 VEGFR-3抗体和 Podoplanin 抗体的脂质靶向微泡,优化制备工艺、流程、脂质配比,免疫荧光评价携载效果,在体实验验证显影效果。结果筛选得出最佳膜材料配比、制备工艺及流程,成功制备出抗体高携载率靶向微泡,平均粒径099μm,平均抗体结合率813%；活体成像研究,靶向微泡可以清晰显示肿瘤新生淋巴管及转移淋巴结；时间-强度曲线证实靶向微泡靶区聚集更多,回声更强,驻留时间更长,实现增强显像。结论优化制备的载 VEGFR-3抗体及 Podoplanin 抗体微泡粒径小而均一性好,抗体装载率高,在体显影效果好,是一种新的具有应用价值的肿瘤新生淋巴管及淋巴结显像剂。

Objective To optimizing preparate for target microbubbles of tumor lymphangiogenesis which load anti-VEGFR-3 and anti-Podoplanin and to evaluate the imaging of this microbubbles Methods The biotinylated anti-VEGFR-3 and anti-Podoplanin were conjugated to biotinylated microbubbles through biotin-avidin its technology of preparation and progress were optimized The proportion of reagents were adjusted The targeted microbubbles were evaluated by immunofluorescence method and its effect was tested in vivo Results High adhesion rate target microbubbles were successfully achieved which average particle size was 0 99 μm and the average antibody combined rate was 81 3% Confirmed by flow cytometry and fluorescence microscope it can clearly show the lymphangiogenesis and the metastasis of lymph nodes in vivo Time-intensity curve showed more microbubbles more intensity more stay and better image Conclusions These target microbubbles have a small particle size and high adhesion rate and a better contrast imaging It is a more value ultrasound target contrast agents of lymph vessel with tumor and sentinel node.