Subtractive SELEX against two heterogeneous target samples: Numerical simulations and analysis
Subtractive SELEX against two heterogeneous target samples: Numerical simulations and analysis
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DOI：
10.1016/j.compbiomed.2006.06.015
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发表时间：
2007-06-01
影响因子：
7.7
通讯作者：
Lin, Lung-Ying
中科院分区：
文献类型：
--
作者：
Chen, Chi-Kan;Kuo, Tzy-Ling;Lin, Lung-Ying
配体指数系统进化（SELEX）是一项革命性的技术，它将组合化学与高通量筛选相结合，从合成的核酸配体文库中生成高亲和力的核酸配体（适体），用于感兴趣的靶标。近年来，SELEX实验已从单一纯化靶标靶向配体文库发展到多个异构靶标样品。这种新应用有可能为药物发现带来巨大的技术和经济优势，但其性能难以预测。为了深入了解新方法，我们开发了一个计算机模型，对两个不同的未知目标异质样本进行数值分析。该模型具有配体文库离散化、配体与靶结合平衡方程离散化、实验中结合配体与非结合配体分离效率离散化等特点。通过计算机模拟，我们研究了在不同的实验条件下，嵌入在未定义目标混合物中的目标适体是如何产生的。我们发现迭代筛选方案基本上能够开发所需的适体。另一方面，靶样结构和分离效率可能共同使筛选动态和结果显著多样化。(C) 2006 Elsevier Ltd版权所有。
Systematic evolution of ligands by exponential (SELEX) is a revolutionary technology that integrates combinatorial chemistry with high throughput screening to generate from synthesized nucleic acid ligand libraries the high affinity nucleic acid ligands (aptamers) for interesting targets. Recently, the SELEX experiments have advanced from targeting the ligand libraries by a single purified target to multiple heterogeneous target samples. Having the potential of bringing enormous technical and economical advantages to drug discovery, the new application suffers from unpredictable performances. To gain an insight of the new method, we develop a computer model to numerically analyze the subtractive SELEX alternatively against two distinct heterogeneous samples of unknown targets. The model features the discretization of ligand library, the ligand-target binding equilibrium equations, and the separation efficiency of bound and unbound ligands in experiments. By computer simulations, we investigate how aptamers for desired targets embedded in undefined target mixtures are generated under different experimental conditions. We find the iterative screening scheme is fundamentally capable of developing desired aptamers. On the other hand, target sample configuration and separation efficiency may all together significantly diversify the screening dynamics and results. (C) 2006 Elsevier Ltd. All rights reserved.