Effect of molecular weight of amine end-modified poly(β-amino ester)s on gene delivery efficiency and toxicity.
Effect of molecular weight of amine end-modified poly(β-amino ester)s on gene delivery efficiency and toxicity.
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DOI：
10.1016/j.biomaterials.2012.01.046
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发表时间：
2012-05
期刊：
影响因子：
14
通讯作者：
Anderson, Daniel G.
中科院分区：
文献类型：
--
作者：
Eltoukhy, Ahmed A.;Siegwart, Daniel J.;Alabi, Christopher A.;Rajan, Jay S.;Langer, Robert;Anderson, Daniel G.
胺末端修饰的聚（β-氨基酯）（PBAE）作为质粒DNA（pDNA）的有效的、可生物降解的聚合物载体引起了人们的兴趣。对于阳离子、不可降解的聚合物，例如聚乙烯亚胺（PEI），聚合物分子量（MW）和分子量分布（MWD）显著影响转染活性和细胞毒性。MW对PBAE的DNA转染活性的影响研究较少。我们采用了两种策略，以获得不同的分子量的胺末端修饰的PBAE。在一种方法中，我们合成了四个胺末端修饰的PBAE，每个在15个不同的单体摩尔比，并观察到中间长度的聚合物介导的HeLa细胞中的最佳DNA转染。这些进料比变体的生物物理表征表明，最佳性能与更高的DNA络合效率和更小的纳米颗粒尺寸有关，但与纳米颗粒电荷无关。在第二种方法中，我们使用制备型尺寸排阻色谱法（SEC）来获得定义明确的单分散聚合物级分。我们观察到，大小分级PBAE的转染活性通常随MW增加，这种趋势与DNA结合效率的增加弱相关。此外，该方法允许分离具有比起始材料更大的转染效力的聚合物级分。对于研究人员与逐步增长聚合合成的基因传递聚合物，我们的数据突出了潜在的广泛用途的制备SEC分离单分散聚合物具有改善的性能。总体而言，这些结果有助于阐明聚合物分子量分布对核酸递送的影响，并为下一代基因治疗材料的合理设计提供了见解。
Amine end-modified poly(ß-amino ester)s (PBAEs) have generated interest as efficient, biodegradable polymeric carriers for plasmid DNA (pDNA). For cationic, non-degradable polymers, such as polyethylenimine (PEI), the polymer molecular weight (MW) and molecular weight distribution (MWD) significantly affect transfection activity and cytotoxicity. The effect of MW on DNA transfection activity for PBAEs has been less well studied. We applied two strategies to obtain amine end-modified PBAEs varying in MW. In one approach, we synthesized four amine end-modified PBAEs with each at 15 different monomer molar ratios, and observed that polymers of intermediate length mediated optimal DNA transfection in HeLa cells. Biophysical characterization of these feed ratio variants suggested that optimal performance was related to higher DNA complexation efficiency and smaller nanoparticle size, but not to nanoparticle charge. In a second approach, we used preparative size exclusion chromatography (SEC) to obtain well-defined, monodisperse polymer fractions. We observed that the transfection activities of size-fractionated PBAEs generally increased with MW, a trend that was weakly associated with an increase in DNA binding efficiency. Furthermore, this approach allowed for the isolation of polymer fractions with greater transfection potency than the starting material. For researchers working with gene delivery polymers synthesized by step-growth polymerization, our data highlight the potentially broad utility of preparative SEC to isolate monodisperse polymers with improved properties. Overall, these results help to elucidate the influence of polymer MWD on nucleic acid delivery and provide insight toward the rational design of next-generation materials for gene therapy.
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影响因子：
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作者：
van de Wetering, P;Cherng, JY;Hennink, WE
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