| 何洁,杨莉,李颖嘉,孙学刚,刁建新,李传刚,宾建平,龚渭冰.以短肽K237为配体的靶向脂质体超声造影剂构建方法[J].中国医学影像技术,2011,27(1):3~7 |
| 以短肽K237为配体的靶向脂质体超声造影剂构建方法 |
| Preparation of liposome ultrasonic contrast agent with ligand peptide K237 |
| 投稿时间：2010-04-07 修订日期：2010-06-29 |
| DOI： |
| 中文关键词: 微泡 流式细胞术 |
| 英文关键词:Microbubbles Flow cytometry |
| 基金项目:国家自然科学基金资助面上项目(30670580)、广东省博士启动基金(9451051501003761)。 |
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| 中文摘要: |
| 目的 探讨以与血管内皮生长因子(VEGF)的主要受体KDR特异性结合的短肽K237(P)为配体制备靶向脂质体超声造影剂(P-Bio-Av-Bio-Mbs)的方法。 方法 采用生物素-亲和素桥接法构建P-Bio-Av-Bio-Mbs,流式细胞术筛选最佳配体适配剂量,光镜及荧光显微镜观察靶向微泡与KDR强阳性表达的人大肠癌LOVO细胞结合情况,计算花环形成率。分别以5、50、99 ml/h速率水流冲刷,光镜观察靶向微泡与LOVO细胞结合情况。 结果 不同亲和素剂量下(0、2、6、10、30 μg),微泡表面亲和素携带率差异有统计学意义(P<0.05)。Mavidin=6 μg时,携带率增长达平台期;不同短肽剂量下(0、30、40、50、60、70、100 μg),微泡表面短肽携带率差异有统计学意义(P<0.05),当MK237=50 μg时,微泡表面短肽携带率增长达平台期。光镜下KDR强阳性表达的LOVO细胞周围花环形成率高达90.52%,荧光显微镜下微泡外壳发出明亮绿色荧光。随冲刷速度增加,靶细胞周围黏附的靶向微泡减少,在99 ml/h冲刷速度下,靶细胞周围仍可见花环结构。 结论 通过生物素-亲和素桥连作用,短肽K237被有效装配在P-Bio-Av-Bio-Mbs表面,体外具有靶向特异性及一定稳定性。流式细胞术是筛选靶向微泡配体适配剂量的可靠方法。 |
| 英文摘要: |
| Objective To assess preparation method of a new kind of targeted liposome ultrasonic contrast agent with small peptide K237 as the ligand which can combine specifically with KDR as the main receptor of VEGF. Methods Targeted bubbles (P-Bio-Av-Bio-Mbs) were formed through "biotin-avidin" bridge grafting. Flow cytometry screening was performed to explore the best dose of the ligands, then targeted-bubbles were incubated respectively with LOVO and LS174T which were KDR expressed in different cells. Meanwhile, rosette formation rate was calculated. Results The bubble surface's avidin-carrying rates were significant different (P<0.05) on different dosages of avidin(0, 2, 6, 10, 30 μg).When Mavidin=6 μg, the avidin labelling ratio reached plateau. There were significant differences in the ratio of peptide K237 labelling (P<0.05) as different peptide dosages (0, 30, 40, 50, 60, 70, 100 μg). When Mk237=50 μg,the peptide labelling ratio reached plateau. In KDR sharply positive expressed LOVO cells, the surrounding rosette formation rate was as high as 90.52% with strong fluorescence intensity. Targeted microbubbles decreased as the water velocity increased. When the velocity reached 99 ml/h, rosette formation still could be seen surrounding the targeted cells. Conclusion KDR-targeted liposome contrast agent with small peptide liganded has been successfully prepared through biotin-avidin mediation, and showed special targeting ability and stability in vitro. Flow cytometry can quantitatively analyze the best dose of ligands carrying targeted microbubbles. |
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