Tombelli S, Minunni M, Mascini M
Università degli Studi di Firenze, Dipartimento di Chimica, Polo Scientifico, Via della Lastruccia 3, 50019 Sesto Fiorentino (FI), Italy.
Biosens Bioelectron. 2005 Jun 15;20(12):2424-34. doi: 10.1016/j.bios.2004.11.006. Epub 2004 Dec 16.
So far, several bio-analytical methods have used nucleic acid probes to detect specific sequences in RNA or DNA targets through hybridisation. More recently, specific nucleic acids, aptamers, selected from random sequence pools, have been shown to bind non-nucleic acid targets, such as small molecules or proteins. The development of in vitro selection and amplification techniques has allowed the identification of specific aptamers, which bind to the target molecules with high affinity. Many small organic molecules with molecular weights from 100 to 10,000 Da have been shown to be good targets for selection. Moreover, aptamers can be selected against difficult target haptens, such as toxins or prions. The selected aptamers can bind to their targets with high affinity and even discriminate between closely related targets. Aptamers can thus be considered as a valid alternative to antibodies or other bio-mimetic receptors, for the development of biosensors and other analytical methods. The production of aptamers is commonly performed by the SELEX (systematic evolution of ligands by exponential enrichment) process, which, starting from large libraries of oligonucleotides, allows the isolation of large amounts of functional nucleic acids by an iterative process of in vitro selection and subsequent amplification through polymerase chain reaction. Aptamers are suitable for applications based on molecular recognition as analytical, diagnostic and therapeutic tools. In this review, the main analytical methods, which have been developed using aptamers, will be discussed together with an overview on the aptamer selection process.
到目前为止,已有多种生物分析方法利用核酸探针通过杂交检测RNA或DNA靶标中的特定序列。最近,从随机序列库中筛选出的特定核酸——适配体,已被证明能与非核酸靶标结合,如小分子或蛋白质。体外筛选和扩增技术的发展使得鉴定出能与靶标分子高亲和力结合的特定适配体成为可能。许多分子量在100至10,000道尔顿之间的小有机分子已被证明是良好的筛选靶标。此外,还可以针对诸如毒素或朊病毒等难以处理的靶标半抗原来筛选适配体。筛选出的适配体能与它们的靶标高亲和力结合,甚至能区分密切相关的靶标。因此,在生物传感器和其他分析方法的开发中,适配体可被视为抗体或其他仿生受体的有效替代品。适配体的制备通常通过SELEX(指数富集的配体系统进化)过程进行,该过程从大量寡核苷酸文库开始,通过体外筛选的迭代过程以及随后的聚合酶链反应扩增,从而分离出大量功能性核酸。适配体适用于基于分子识别的分析、诊断和治疗工具等应用。在本综述中,将讨论使用适配体开发的主要分析方法,并概述适配体的筛选过程。