摘要
目的制备携抗P-选择素（P-seleetin）单抗的靶向超声造影剂，并研究其体外寻靶能力。方法采用“亲和素-生物素”桥接法制备携抗P—selectin单抗靶向超声造影剂，检测其基本特性。应用不同剂量重组人白介素4（rhIL-4）和组胺联合刺激人脐静脉血管内皮细胞（HuVEcs）表达P—selectin，用免疫荧光检测P—selectin的表达水平并探寻rhIL-4最适刺激剂量。靶向超声造影剂、同型对照超声造影剂和空白超声造影剂分别与经最适剂量rhIL-4和组胺联合刺激的HUVECs和未作处理的HUVECs进行体外寻靶实验。结果成功制备携抗P—selectin单抗靶向超声造影剂，平均粒径为（2．24±0．71）弘m，平均Zeta电位为（-2．75±0．84）mV，浓度为（3．0±0．3）×10^9个／ml，抗体结合率高达99．80％；免疫荧光检测结果显示rhIL-4和组胺能成功刺激HUVECs表达P—selectin，并探寻出rhlL一4最适刺激剂量；体外寻靶实验显示靶向超声造影剂能较好地黏附在经最适剂量rhIL-4和组胺联合刺激的HUVECs表面，与其他组比较差异有统计学意义（P〈0．01）。结论采用“亲和素一生物素”桥接法成功制备携抗P—selectin单抗靶向超声造影剂，该造影剂在体外能有效地黏附在经rhIL-4和组胺联合刺激的HUVECs表面。
Objective To prepare a kind of P-selectin antibody loaded-targeted ultrasound contrast agent,and to investigate its ability of targeting in vitro. Methods P-selectin antibody loaded-targeted ultrasound contrast agent was prepared via an ＂avidin-biotin＂ bridge. Its basic property was determined. Human umbilical vein endothelial cells （HUVECs） were stimulated to express P-selectin by applying different doses of recombinant human interleukin-4 （rhIL-4） and histamine. The expression level of P-selectin was detected by immunofluorescence technique, and explored rhlL-4 optimal stimulation dose. The experiment group was divided into 3 groups, including targeted ultrasound contrast agent, isotype control ultrasound contrast agent and blank ultrasound contrast agent. Targeting abilities of 3 groups were observed by adhering to treated HUVECs and untreated HUVECs,respectively. Results P-selectin antibody loaded- targeted ultrasound contrast agent was prepared successfully. The average diameter was （2.24± 0.71）μm. The average Zeta potential was （ - 2.75 ± 0.84） inV. The concentration was （3.0 ±0.3） × 10^9/ml. The rate of antibody binding was as high as 99.80%. RhIL-4 and histamine could stimulate HUVECs to express P- selectin,and optimal stimulation dose was obtained. Targeting experiment showed that targeted ultrasound contrast agent could preferably adhere to HUVECs stimulated by optimal dose of rhIL-4 and histamine. Compared with other groups, there were significant differences. Conclusions P-selectin antibody-loaded targeted ultrasound contrast agent was prepared successfully via an ＂avidin-biotin＂ bridge. The targeted ultrasound contrast agent could effectively adhere to HUVECs stimulated by rhIL-4 and histamine.
出处
《中华超声影像学杂志》
CSCD
北大核心
2013年第4期349-354,共6页
Chinese Journal of Ultrasonography
基金
国家自然科学基金青年科学基金项目（81000621）
教育部博士点（新教师类）基金项目（20105503120008）
国家自然科学基金重点项目（81130025）
2010年度重庆高校优秀成果转化资助项目（Kjzh10201）
