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利用层层自组装法制备功能性适配体薄膜。

Preparation of functional aptamer films using layer-by-layer self-assembly.

作者信息

Sultan Yasir, Walsh Ryan, Monreal Carlos, DeRosa Maria C

机构信息

Department of Chemistry, Carleton University, Ottawa-Carleton Chemistry Institute, 1125 Colonel By Drive, Ottawa, Ontario, Canada K1S 5B6.

出版信息

Biomacromolecules. 2009 May 11;10(5):1149-54. doi: 10.1021/bm8014126.

Abstract

Advances in many aptamer-based applications will require a better understanding of how an aptamer's molecular recognition ability is affected by its incorporation into a suitable matrix. In this study, we investigated whether a model aptamer system, the sulforhodamine B aptamer, would retain its binding ability while embedded in a multilayer polyelectrolyte film. Thin films consisting of poly(diallyldimethylammonium chloride) as the polycation and both poly(sodium 4-styrene-sulfonate) and the aptamer as the polyanions were deposited by the layer-by-layer approach and were compared to films prepared using calf thymus DNA or a random single-stranded oligonucleotide. Data from UV-vis spectroscopy, quartz crystal microbalance studies, confocal microscopy, and time of flight secondary ion mass spectrometry confirm that the aptamer's recognition of its target is retained, with no loss of specificity and only a modest reduction of binding affinity, while it is incorporated within the thin film. These findings open up a raft of new opportunities for the development and application of aptamer-based functional thin films.

摘要

许多基于适配体的应用取得进展将需要更好地理解适配体的分子识别能力如何受到其掺入合适基质的影响。在本研究中,我们调查了一种模型适配体系统——磺基罗丹明B适配体,在嵌入多层聚电解质膜时是否会保留其结合能力。由聚(二烯丙基二甲基氯化铵)作为聚阳离子以及聚(4-苯乙烯磺酸钠)和适配体作为聚阴离子组成的薄膜通过逐层方法沉积,并与使用小牛胸腺DNA或随机单链寡核苷酸制备的薄膜进行比较。紫外可见光谱、石英晶体微天平研究、共聚焦显微镜和飞行时间二次离子质谱的数据证实,适配体对其靶标的识别得以保留,特异性没有丧失,结合亲和力仅适度降低,同时它被掺入薄膜中。这些发现为基于适配体的功能薄膜的开发和应用开辟了一系列新机会。

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