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不同壳层包覆的磁性纳米颗粒用于生物识别:高特异性结合能力。

Magnetic nanoparticles coated with different shells for biorecognition: high specific binding capacity.

机构信息

Gazi University, Faculty of Science, Department of Chemistry, 06500 Besevler, Ankara, Turkey.

出版信息

Analyst. 2014 Mar 7;139(5):1093-100. doi: 10.1039/c3an01726e. Epub 2014 Jan 10.

Abstract

Modifying the surfaces of magnetic nanoparticles (MNPs) by the covalent attachment of biomolecules will enable their application as media for magnetically-assisted bioseparations. In this paper, we reported both the activity and specific binding capacity of ferritin antibodies on core-shell MNPs. The antibodies were covalently attached on silica-, silver- and polydopamine-coated MNPs by different methods. Anti-ferritin was bound onto the silica- or silver-coated MNPs by conventional methods using 3-aminopropyltriethoxysilane (APTES) or 11-mercaptoundecanoic acid (MUA), which was followed by activation of carboxyl groups by EDC/NHS. However with anti-ferritin immobilized onto the Fe3O4 nanoparticles modified with polydopamine, an in situ coating formed through the adhesive proteins. In addition, a great deal of anti-ferritin biomolecules covalently attached onto the MNPs. According to our results, the amounts of bound anti-ferritin onto the silica-, silver- and PDA-coated MNPs were 70, 75 and 95 μg anti-ferritin per mg MNP, respectively. In the experiments, polydopamine (PDA)-coated MNPs showed faster adsorption, more significant selectivity and a larger binding capacity than the others. Also, the equilibrium dissociation constants of the antigen-antibody complexes were determined on the anti-ferritin-immobilized MNPs. Silica-, PDA- and silver-coated MNPs had Kd values of 5.45 × 10(-7), 2.12 × 10(-7) and 3.91 × 10(-8) mol L(-1), respectively. Based on these results, the affinity of the anti-ferritin for ferritin on the PDA-coated MNPs was approximately 10-fold higher than that on the silica- and silver-coated MNPs. In addition, among the anti-ferritin-immobilized silica-, silver- and PDA-coated MNPs, the PDA-coated MNPs showed the highest antigen selectivity values. As a result, anti-ferritin-immobilized PDA-coated MNPs represented a higher activity and stronger affinity for the specific antigen than the others.

摘要

通过共价键将生物分子附着在磁性纳米粒子 (MNPs) 的表面上,可以使它们能够作为磁辅助生物分离的介质。在本文中,我们报告了铁蛋白抗体在核壳 MNPs 上的活性和特异性结合能力。通过不同的方法将抗体通过共价键附着在二氧化硅、银和聚多巴胺涂层的 MNPs 上。使用 3-氨基丙基三乙氧基硅烷 (APTES) 或 11-巯基十一酸 (MUA) 通过常规方法将抗铁蛋白结合到二氧化硅或银涂层的 MNPs 上,随后通过 EDC/NHS 激活羧基。然而,与通过聚多巴胺修饰的 Fe3O4 纳米粒子固定的抗铁蛋白一起,通过粘合蛋白形成了原位涂层。此外,大量的抗铁蛋白生物分子共价附着在 MNPs 上。根据我们的结果,结合到二氧化硅、银和 PDA 涂层 MNPs 上的结合抗铁蛋白的量分别为 70、75 和 95 μg 抗铁蛋白/毫克 MNP。在实验中,与其他方法相比,聚多巴胺 (PDA) 涂层 MNPs 表现出更快的吸附速度、更显著的选择性和更大的结合能力。此外,还在固定有抗铁蛋白的 MNPs 上确定了抗原-抗体复合物的平衡解离常数。二氧化硅、PDA 和银涂层 MNPs 的 Kd 值分别为 5.45×10(-7)、2.12×10(-7) 和 3.91×10(-8)mol·L(-1)。基于这些结果,抗铁蛋白与 PDA 涂层 MNPs 上的铁蛋白的亲和力大约比二氧化硅和银涂层 MNPs 高 10 倍。此外,在固定有抗铁蛋白的二氧化硅、银和 PDA 涂层 MNPs 中,PDA 涂层 MNPs 表现出最高的抗原选择性值。因此,与其他方法相比,固定有抗铁蛋白的 PDA 涂层 MNPs 对特定抗原表现出更高的活性和更强的亲和力。

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