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高通量传递和光谱差异对生物分子纳米缀合物的表征。

Characterization of biomolecular nanoconjugates by high-throughput delivery and spectroscopic difference.

机构信息

Missouri State University, Cell & Molecular Biology Program, Springfield, MO 65897, USA.

出版信息

Nanomedicine (Lond). 2012 Dec;7(12):1851-62. doi: 10.2217/nnm.12.70. Epub 2012 Sep 3.

Abstract

AIM

Nanoparticle conjugates have the potential for delivering siRNA, splice-shifting oligomers or nucleic acid vaccines, and can be applicable to anticancer therapeutics. This article compares tripartite conjugates with gold nanoparticles or synthetic methoxypoly(ethylene glycol)-block-polyamidoamine dendrimers.

MATERIALS & METHODS: Interactions with model liposomes of a 1:1 molar ratio of tripalmitin:cholesterol or phospholipid:cholesterol were investigated by high-throughput absorbance, as well as fluorescence difference and cellular luminescence assays.

RESULTS

Spectral differences and dynamic light-scattering spectroscopy shifts demonstrated the interaction of conjugates with liposomes. Biological activity was demonstrated by upregulation of gene expression via splice-shifting oligomers, delivery of anti-B-Raf siRNA in cultured human cancer cells or tuberculosis antigen 85B plasmid expression vector in a coculture model of antigen presentation.

CONCLUSION

The data suggests that gold nanoparticles and methoxypoly(ethylene glycol)-block-polyamidoamine dendrimer nanoconjugates may have potential for binding, stabilization and delivery of splice-shifting oligomers, siRNA and nucleic acid vaccines for preclinical trials.

摘要

目的

纳米粒子缀合物具有递送 siRNA、剪接移位寡聚物或核酸疫苗的潜力,并且可适用于抗癌治疗。本文比较了三联缀合物与金纳米粒子或合成甲氧基聚(乙二醇)-嵌段-聚酰胺胺树枝状大分子。

材料与方法

通过高通量吸光度法以及荧光差示和细胞发光测定法,研究了三棕榈酸甘油酯:胆固醇或磷脂:胆固醇摩尔比为 1:1 的模型脂质体与三部分缀合物的相互作用。

结果

光谱差异和动态光散射光谱的移动表明了缀合物与脂质体的相互作用。通过剪接移位寡聚物上调基因表达、在培养的人类癌细胞中递送抗 B-Raf siRNA 或在抗原呈递的共培养模型中表达结核抗原 85B 质粒表达载体,证明了生物活性。

结论

数据表明,金纳米粒子和甲氧基聚(乙二醇)-嵌段-聚酰胺胺树枝状大分子纳米缀合物可能具有结合、稳定和递送电镜移位寡聚物、siRNA 和核酸疫苗的潜力,可用于临床前试验。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7bcb/4295771/d1b22d353186/nihms-432614-f0001.jpg

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