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通过主客体相互作用在硅纳米颗粒表面接枝聚丙烯酸刷层以用于检测乙型肝炎病毒的 DNA 序列

Silica nanoparticles coated by poly(acrylic acid) brushes via host-guest interactions for detecting DNA sequence of Hepatitis B virus.

机构信息

Lab of Advanced Materials, State Key Laboratory of Molecular Engineering of Polymers, Collaborative Innovation Center of Chemistry for Energy Materials, Fudan University, Shanghai 200438, PR China.

Lab of Advanced Materials, State Key Laboratory of Molecular Engineering of Polymers, Collaborative Innovation Center of Chemistry for Energy Materials, Fudan University, Shanghai 200438, PR China.

出版信息

Talanta. 2018 May 1;181:65-72. doi: 10.1016/j.talanta.2017.12.061. Epub 2017 Dec 22.

Abstract

Poly(acrylic acid) (PAA) brushes coated onto silica nanoparticles have been widely utilized in bioassays due to their abilities of providing favorable microenvironments and ensuring good biological activities for biomolecules. However, traditional PAA brushes are synthesized by reversible addition-fragmentation chain transfer polymerization. Hence, it is generally difficult to control and characterize the molecular weight of the PAA brushes, which may depress the reproducibility and bring more uncertain results. Herein, atom transfer radical polymerization method is employed to synthesize β-cyclodextrin-cored PAA with uniform and controllable molecular weight. After loading on the surfaces of adamantane-functionalized silica nanoparticles via host-guest interactions, glucose oxidase and probe single strand DNA (ssDNA) are further immobilized on the as-prepared nanoparticles. Meanwhile, capture ssDNA is functionalized on amino modified magnetic beads. In the presence of ssDNA sequence of Hepatitis B Virus (HBV) containing completely matched sequence of both probe and capture ssDNA, a bioconjugate is formed and can be separated by an external magnet. The isolated glucose oxidase can further catalyze glucose into gluconic acid and HO, and then reduce HAuCl on Au seeds. By monitoring the absorption intensity change of the Au NPs at 530nm, the proposed biosensor with novel signal amplification probes can be used to detect DNA sequence of HBV with high sensitivity and selectivity in both buffer and serum samples. This developed strategy has presented a new way to construct silica nanoparticles coated by PAA brushes for the fields of clinical diagnosis and other life sciences.

摘要

聚(丙烯酸)(PAA)刷涂在硅胶纳米粒子上,由于其能够提供有利的微环境并确保生物分子具有良好的生物活性,因此已广泛应用于生物测定中。然而,传统的 PAA 刷是通过可逆加成-断裂链转移聚合合成的。因此,通常难以控制和表征 PAA 刷的分子量,这可能会降低重现性并带来更多不确定的结果。在此,采用原子转移自由基聚合方法合成了具有均匀可控分子量的β-环糊精核 PAA。通过主客体相互作用将其负载在金刚烷功能化硅胶纳米粒子表面后,进一步将葡萄糖氧化酶和探针单链 DNA(ssDNA)固定在制备的纳米粒子上。同时,将捕获 ssDNA 功能化在氨基修饰的磁性珠上。在含有与探针和捕获 ssDNA 完全匹配序列的乙型肝炎病毒(HBV)ssDNA 序列存在下,形成生物缀合物,并可通过外部磁场分离。分离的葡萄糖氧化酶可进一步将葡萄糖催化成葡萄糖酸和 HO,然后在 Au 种子上还原 HAuCl。通过监测 Au NPs 在 530nm 处的吸收强度变化,可以使用具有新型信号放大探针的这种生物传感器在缓冲液和血清样品中以高灵敏度和选择性检测 HBV 的 DNA 序列。该方法为临床诊断和其他生命科学领域中 PAA 刷涂硅胶纳米粒子的构建提供了一种新方法。

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