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Sequence-constructive SELEX: A new strategy for screening DNA aptamer binding to Globo H

机译:序列构建性SELEX:筛选DNA适体与Globo H结合的新策略

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摘要

We proposed to use a novel stepwise sequence-constructive SELEX method to develop DNA aptamers that can recognize Globo H which is a tumor-associated carbohydrate antigen. A combinatorial synthetic library that consisted of DNA molecules with randomized regions of 15-bases was used as the starting library for the first SELEX procedure. The input DNA library for the second round of SELEX consisted of the extension of the 5' and 3'-ends with 7-bases that were randomized from four selected aptamers. The third round of SELEX was performed following the same procedures as described for the second round of SELEX. The experimental results indicate that the binding affinity of DNA aptamers to Globo H was enhanced when using the sequence-constructive SELEX approach. The selectivity of the DNA aptamers for related disaccharides, mannose derivatives, and Globo H analogs demonstrated the ability of the DNA aptamers to discriminate the presence of various glycans with different structures. (C) 2014 Elsevier Inc. All rights reserved.
机译:我们建议使用一种新颖的逐步构建序列的SELEX方法来开发可以识别Globo H的DNA适体,Globo H是一种与肿瘤相关的碳水化合物抗原。包含15个碱基随机区域的DNA分子组成的组合合成库用作第一个SELEX程序的起始库。第二轮SELEX的输入DNA文库由5个末端和3个末端的7个碱基组成,这些碱基从4个选定的适体中随机分配。按照与第二轮SELEX相同的程序执行第三轮SELEX。实验结果表明,当使用序列构建SELEX方法时,DNA适体与Globo H的结合亲和力得到了增强。 DNA适体对相关二糖,甘露糖衍生物和Globo H类似物的选择性证明了DNA适体区分各种具有不同结构的聚糖的能力。 (C)2014 Elsevier Inc.保留所有权利。

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