血栓靶向脂膜超声造影剂的制备与初步评价

Preparation of thrombus-targeted lipid-coated ultrasound contrast agent and initial assessment

目的制备血栓靶向脂膜超声造影剂,并对其配体结合率进行初步评价。方法通过生物素-抗生物素蛋白桥连法,制备携带血栓靶向配体的脂膜超声造影剂,非靶向脂膜微泡为对照;采用流式细胞仪初步评价靶向微泡配体结合率及其影响因素。结果靶向微泡荧光检测为阳性,对照组为阴性;流式细胞仪分析结果显示该血栓靶向微泡配体结合率达82.96%,对照组仅为0.92%、0.89%。结论采用生物素-抗生物素蛋白桥连接法,可以成功制备血栓靶向脂膜超声造影剂。微泡纯化过程中离心速度与离心时间对配体结合率的影响具有显著意义。

Objective To prepare thrombus-specific targeted lipid-coated microbubbles and investigate their properties. Methods The binding is acquired by biotin-avidin conjugation. Immunofluorescence was applied to confirm the conjugation. The conjunct ratio was accessed by flow cytometer （FCM）. Non-targeted microbubbles was used as the control. Results The targeted microbubbles conjugated with biotin-avidin were positive and gave out shining bright fluorescence in immunofluorescent assay. While in the control group, the lack of fluorescence demonstrared a poor bubbles specific binding. FCM demonstrate that the shell＇s wavelength of thrombus-targeted microbubbles changed greatly, and the conjunct ratios reached 82.96%, while control groups were 0.92% and 0.89%. Conclusion Thrombus-targeted lipid-coated microbubbles with high specific biological activity were prepared successfully by biotin-avidin conjugation. The effect of centrifuge velocity and time was significant during microbubbles sublimation.