超声微泡连接特异性抗体的制备方法及靶向化处理因素对超声微泡物理性质的影响

目的 探讨超声微泡连接特异性抗体的制备方法及靶向化处理因素对超声微泡物理性质的影响.方法 应用生物素-链霉亲和素连接系统,使注射用六氟化硫微泡(SonoVue)与特异性抗血管细胞黏附分子-1(VCAM-1)抗体相连接,用间接免疫荧光法检测抗体与微泡的连接,用马尔文激光粒径分析仪分别测定生物素化处理前后微泡的粒径,采用超声诊断仪评价微泡靶向化构建前后的显像效果.结果 SonoVue与特异性抗体成功连接,间接免疫荧光检测呈阳性.生物素修饰后的微泡粒径分布变窄,与普通微泡的超声显像效果略有差异.结论 通过生物素-链霉亲和素系统可以成功靶向化构建超声微泡,靶向化处理因素对微泡的粒径有一定影响,对微泡的超声显影效果略有影响.

Preparation of targeted antibody connecting to microbubbles and evaluation on the impact of targeting on the physical properties of microbubbles

Objective To investigate the preparing methods of targeted antibody connecting to microbubbles, and to evaluate the impact of targeting on the physical properties of microbubbles. Methods The vascular cell adhesion molecular 1 (VCAM-1) monoclonal antibody was applied to connect microbubbles via biotin-streptavidin system, and the targeted microbubbles were evaluated by using indirect immunofluorenscence. The particle diameters of SonoVue microbubbles and targeted to VCAM-1 microbubbles were tested by using Malvern ZEN 3690. The ultrasonic enhancement of the microbubbles was observed by using ultrasonic diagnostic apparatus. Results SonoVue was successfully connected to targeted antibody. Indirect immunofluorenscence showed that the targeted microbubbles were positive. The diameter distribution of the targeted biotin-modified microbubbles was narrower than of normal SonoVue. There was little difference in ultrasonic enhancement between the targeted SonoVue and normal SonoVue. Conclusion The targeted ultrasound microbubbles is successfully prepared via biotin-streptavidin-biotin system. The processing factors affect on the diameter of the microbubbles, resulting little difference in ultrasonic enhancement.