CD11c免疫脂质体的制备及其体外树突状细胞靶向性
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上海高校选拔培养优秀青年教师科研专项基金(egd09014),第二军医大学专项基金(12Js07).


Preparation of CD11c modified immunoliposome and validation of its targeting to dendritic cells
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Supported by Special Funds of Shanghai for Cultivation and Selection of Excellent Young Teachers for Higher Education (egd09014) and Special Fund of Second Military Medical University (12Js07).

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    摘要:

    目的 制备CD11c单抗修饰的免疫脂质体并进行表征,验证其体外对树突状细胞(dendritic cells,DCs)的特异靶向性。方法 采用薄膜分散法制备脂质体,通过化学交联法将CD11c单抗连接于脂质体上,利用高效液相法测定CD11c与脂质体的连接率,透射电镜观察脂质体形态,激光粒度测定仪测定粒径和Zeta电势,流式细胞仪检测脂质体上连接CD11c的活性和对DCs的靶向效果,激光共聚焦显微镜观察该免疫脂质体特异性靶向特点。 结果 成功制备了CD11c免疫脂质体,测得单抗连接率为(77.6±3.2)%,平均粒径为(151.2±1.6) nm,Zeta电位为(-32.0±1.8) mV。电镜下脂质体粒径均匀,表面光滑,多为圆形,少许椭圆形,无粘连。CD11c单抗连接于脂质体仍保持了良好的活性,体外表现出显著的DCs特异靶向性。结论 制备的CD11c免疫脂质体对DCs细胞具有特异靶向性。

    Abstract:

    Objective To prepare and characterize liposome with mAb(CD11c-imliposomes) and to validate its targeting to dendritic cells (DCs) in vitro. Methods Liposomes were prepared by film dispersion method, and CD11c mAb was collected to the surface of liposome by chemical cross-linking method. The conjugation efficiency of CD11c mAb to the liposome was determined by HPLC; the morphology of CD11c-imliposomes was observed under transmission electron microscope; the particle size and Zata potential were determined by dynamic light-scattering; the activity of CD11c mAb was examined by flow cytometry; and its specific targeting to DCs was visualized by laser scanning confocal microscopy. Results The CD11c-imliposomes were successfully prepared and the conjugation efficiency was (77.6±3.2)%, with the particle size being (151.2±1.6) nm and the Zeta potential being (-32.0±1.8) mV. Under microscope, the CD11c-imliposomes were mostly round in shape or elliptical, with smooth surface and with no adhesion. The activity of CD11c-imliposomes was still satisfactory and demonstrated great targeting to DCs in vitro. Conclusion The CD11c-imliposomes prepared by us has targeting ability to DCs in vitro.

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  • 收稿日期:2013-01-06
  • 最后修改日期:2013-03-29
  • 录用日期:2013-05-09
  • 在线发布日期: 2013-06-24
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