New strategies for evaluation and analysis of SELEX experiments.

New strategies for evaluation and analysis of SELEX experiments.
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DOI:
10.1155/2014/849743
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发表时间:
2014
影响因子:
--
通讯作者:
Labudde D
Labudde D
中科院分区:
生物学3区
文献类型:
--
作者:
Beier R;Boschke E;Labudde D

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适配子是制药和生物传感器中抗体的一种有趣的替代品,因为它们能够以高亲和力与多种可能的靶分子结合。因此,寻找这种适配子的过程--通常是SELEX筛选过程--变得至关重要。标准的SELEX程序通过结合实验安排对某些已发现的适体进行验证,这不会导致在筛选过程中对适体浓缩进行任何详细的说明。为了进一步分析SELEX文库中积累的富集物,我们除了使用标准的SELEX程序外,还使用了通过下一代测序技术收集的序列信息。由于序列基序是富集描述的一种可能性,因此需要找到与适体内的亚结构相对应的重复序列基序,这些亚结构特征地适合于靶的特定结合位点。本文提出了一种基序搜索算法,该算法有助于更详细地描述适配子的富集化。对靶标和结合适配子的广泛表征稍后可能揭示这些分子之间的功能联系,这可以被建模并用于优化未来的SELEX运行,以防产生靶标特异的起始库。
Aptamers are an interesting alternative to antibodies in pharmaceutics and biosensorics, because they are able to bind to a multitude of possible target molecules with high affinity. Therefore the process of finding such aptamers, which is commonly a SELEX screening process, becomes crucial. The standard SELEX procedure schedules the validation of certain found aptamers via binding experiments, which is not leading to any detailed specification of the aptamer enrichment during the screening. For the purpose of advanced analysis of the accrued enrichment within the SELEX library we used sequence information gathered by next generation sequencing techniques in addition to the standard SELEX procedure. As sequence motifs are one possibility of enrichment description, the need of finding those recurring sequence motifs corresponding to substructures within the aptamers, which are characteristically fitted to specific binding sites of the target, arises. In this paper a motif search algorithm is presented, which helps to describe the aptamers enrichment in more detail. The extensive characterization of target and binding aptamers may later reveal a functional connection between these molecules, which can be modeled and used to optimize future SELEX runs in case of the generation of target-specific starting libraries.
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