Selective targeting of antibody-conjugated nanoparticles to leukemic cells and primary T-lymphocytes
Selective targeting of antibody-conjugated nanoparticles to leukemic cells and primary T-lymphocytes
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DOI：
10.1016/j.biomaterials.2005.02.038
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发表时间：
2005-10-01
期刊：
影响因子：
14
通讯作者：
von Briesen, H
中科院分区：
文献类型：
--
作者：
Dinauer, N;Balthasar, S;von Briesen, H
本研究将基于生物可降解材料的表面修饰纳米粒子用于抗体偶联，以获得选择性的药物载体体系。采用解溶法制备了明胶纳米颗粒。引入了能够连接中性亲和素(NAV)的巯基。针对淋巴细胞表面CD3抗原的抗体被偶联到纳米颗粒表面。通过NAV-生物素-复合体的形成，实现生物素化的抗CD3抗体的结合。用流式细胞仪和激光共聚焦扫描显微镜(CLSM)检测细胞结合和摄取。可以显示抗CD3结合的纳米颗粒针对CD3阳性的人T细胞白血病细胞和原代T淋巴细胞的细胞类型特异性靶向。细胞摄取和有效的内化抗体偶联纳米颗粒CD3表达细胞。T细胞白血病细胞的摄取率约为84%。为了证实T细胞靶向的选择性，在纳米颗粒孵育前加入过量的游离抗CD3抗体进行了竞争实验，导致细胞对抗体偶联纳米颗粒的摄取显著减少。对摄取机制的进一步分析证实了受体介导的内吞过程。基于蛋白质的纳米颗粒与特定细胞抗原的抗体结合，有望成为特定细胞类型的选择性药物输送系统。(C)2005爱思唯尔有限公司。保留所有权利。
In the present study, surface-modified nanoparticles based on biodegradable material were used for antibody coupling in order to get a selective drug carrier systems. Gelatin nanoparticles were prepared by a desolvation process. Sulfhydryl groups were introduced which enabled the linkage of NeutrAvidin (NAv). Antibodies specific for the CD3 antigen on lymphocytic cells were conjugated to the nanoparticles surface. The binding of biotinylated anti-CD3 antibody was achieved by NAv-biotin-complex formation. Cellular binding and uptake were determined by flow cytometry and confocal laser scanning microscopy (CLSM). Celltype-specific targeting of anti-CD3-conjugated nanoparticles into CD3-positive human T-cell leukemia cells and primary T-lymphocytes could be shown. Celluar uptake and effective internalization of antibody-conjugated nanoparticles into CD3 expressing cells were demonstrated. Uptake rates of about 84% into T-cell leukemia cells were observed. To confirm selectivity of T-cell targeting, competition experiments were carried out adding excessive free anti-CD3 prior to nanoparticle incubation leading to significantly reduced cellular uptake of antibody-conjugated nanoparticles. Further analysis on the mechanism of uptake confirmed a receptor-mediated endocytotic process. Protein-based nanoparticles conjugated with an antibody against a specific cellular antigen hold promise as selective drug delivery systems for specific cell types. (c) 2005 Elsevier Ltd. All rights reserved.