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

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.