Electrophoretic purification of tumor-targeted polyethylenimine-based polyplexes reduces toxic side effects in vivo
Electrophoretic purification of tumor-targeted polyethylenimine-based polyplexes reduces toxic side effects in vivo
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DOI：
10.1016/j.jconrel.2007.05.013
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发表时间：
2007-10-08
影响因子：
10.8
通讯作者：
Ogris, Manfred
中科院分区：
文献类型：
--
作者：
Fahrmeir, Julia;Gunther, Michael;Ogris, Manfred
基于聚乙烯亚胺 (PEI) 的非病毒载体通常使用过量的 PEI 生成。然而，未结合的聚合物的量与限制这些基因载体的体内使用的毒性相关。基于小于 200 nm 的 PEI/DNA 复合物的尺寸排阻色谱法的纯化已被证明可以有效去除未结合的 PEI 聚合物。一种基于电泳的新型纯化方法可以纯化 PEI 聚合复合物，而不受其大小的影响，从而产生最终 PEI 氮/DNA 磷酸盐比率在 2.6 至 3.1 之间的聚合复合物。未结合的 PEI 缀合物（例如聚乙二醇化 PEI 和转铁蛋白缀合 PEI）也可以从复合物中分离出来，从而提供具有明确成分的制剂。纯化的复合物可以以高转染效率介导体外基因转移，同时表现出较低的细胞毒性。当以100μg/20g体重全身递送至荷瘤小鼠时，纯化的聚合复合物具有良好的耐受性，肿瘤基因表达水平比未纯化的聚合复合物高5倍。接受未纯化基因载体的小鼠表现出严重的毒性，导致高死亡率和不利的基因表达模式。 mu 2007 Elsevier B.V. 保留所有权利。
Non-viral vectors based on polyethylenimine (PEI) are usually generated with an excess of PEI. However, the amount of unbound polymer correlates with toxicity limiting the in vivo use of these gene carriers. Purification based on size exclusion chromatography of PEI/DNA polyplexes smaller than 200 nm has been shown to efficiently remove unbound PEI polymer. A novel purification method based on electrophoresis can purify PEI polyplexes independent of their size resulting in polyplexes with final PEI nitrogen/DNA phosphate ratios between 2.6 and 3.1. Also unbound PEI conjugates like PEGylated PEI and transferrin-conjugated PEI can be separated from the polyplexes, providing formulations with clearly defined compositions. Purified polyplexes can mediate in vitro gene transfer with high transfection efficiencies while demonstrating lower cellular toxicity. Purified polyplexes were well-tolerated when systemically delivered into tumor-bearing mice at 100 mu g/20 g body weight, with tumor gene expression levels up to 5-fold higher than the non-purified polyplexes. Mice receiving non-purified gene carriers exhibited severe toxicity leading to high mortality and unfavourable gene expression patterns. mu 2007 Elsevier B.V. All rights reserved.