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聚(乙烯亚胺)接枝聚[(天冬氨酸)-共-赖氨酸],一种潜在的用于DNA递送的非病毒载体。

Poly (ethylenimine)-grafted-poly [(aspartic acid)-co-lysine], a potential non-viral vector for DNA delivery.

作者信息

Tang Gu Ping, Yang Zhi, Zhou Jun

机构信息

Institute of Bioengineering and Nanotechnology, Singapore.

出版信息

J Biomater Sci Polym Ed. 2006;17(4):461-80. doi: 10.1163/156856206776374133.

Abstract

A potential non-viral gene-transfer vector, poly(ethylenimine)-grafted-poly[(aspartic acid)-co-lysine] (PSL), has been developed by thermal polycondensation of aspartic acid and lysine under reduced pressure. Low-molecular-mass branch poly(ethylenimine) (PEI600) was conjugated to the backbone. The chemical structure of the resulting co-polymer was identified by 1H-NMR, FT-IR, TGA and X-ray diffraction. The results of the MTT assay showed that at concentration up to 4000 nmol/l of the vector cell viability was over 80% and showed low toxicity. Electrophoretic retardation and ethidum bromide assay showed that at N/P ratios 12-15 (w/w) the DNA could be condensed and neutralized. Using the zeta potential assay we discovered that it had a high positive charge on its surface of the particle (over 30 mV). The particle sizes of the co-polymer/DNA complexes were 150-170 nm, as measured by DLS and AFM. Compared with PEI600, co-polymer/DNA complexes showed a significant enhancement of transfection activity in the absence and presence of serum in NT2 and COS7 cell lines. This means that the PEI600-PSL co-polymer is a promising candidate for gene delivery.

摘要

一种潜在的非病毒基因传递载体,聚(乙烯亚胺)接枝聚[(天冬氨酸)-共-赖氨酸](PSL),已通过天冬氨酸和赖氨酸在减压下的热缩聚反应制备而成。低分子量支链聚(乙烯亚胺)(PEI600)与主链相连。通过1H-NMR、FT-IR、TGA和X射线衍射对所得共聚物的化学结构进行了鉴定。MTT试验结果表明,在载体浓度高达4000 nmol/l时,细胞活力超过80%,且毒性较低。电泳阻滞和溴化乙锭试验表明,在N/P比为12-15(w/w)时,DNA能够被凝聚和中和。通过zeta电位测定法,我们发现该颗粒表面带有高正电荷(超过30 mV)。通过动态光散射(DLS)和原子力显微镜(AFM)测量,共聚物/DNA复合物的粒径为150-170 nm。与PEI600相比,在NT2和COS7细胞系中,无论有无血清存在,共聚物/DNA复合物的转染活性均显著增强。这意味着PEI600-PSL共聚物是一种很有前景的基因传递候选物。

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