SELMA: Selection with Modified Aptamers.
SELMA: Selection with Modified Aptamers.
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DOI：
10.1002/9780470559277.ch140233
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发表时间：
2015-06-01
影响因子：
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通讯作者：
Krauss IJ
中科院分区：
文献类型：
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作者：
Temme JS;Krauss IJ
核酸适体的体外选择，创造了SELEX，导致了新的治疗方法的发现，并有助于许多配体-生物分子相互作用的结构和机制的理解。一种相关的方法，用修饰的适体选择（SELMA），使得能够选择含有不能进行PCR的具有大修饰的碱基的DNA适体。这种方法的一个关键应用是含有碳水化合物修饰的适体的进化。碳水化合物结合蛋白通常需要几个拷贝的碳水化合物部分以进行强识别。尽管可能难以合理地设计以用于靶识别的最佳间距和方向聚集聚糖的合成支架，但SELMA使糖适体具有高度优化的聚糖聚集，从而实现低纳摩尔识别。尽管可以设想许多应用，但本文中的方案和讨论描述了将SELMA应用于发现与HIV广泛中和抗体2G 12结合的glycoDNAs所涉及的程序。
In vitro selection of nucleic acid aptamers, coined SELEX, has led to the discovery of novel therapeutics and aided in the structural and mechanistic understanding of many ligand-biomolecule interactions. A related method, selection with modified aptamers (SELMA), enables selection of DNA aptamers containing bases with a large modification that cannot undergo PCR. A key application of this method is the evolution of aptamers containing carbohydrate modifications. Carbohydrate-binding proteins normally require several copies of the carbohydrate moiety for strong recognition. Whereas it may be difficult to rationally design synthetic scaffolds that cluster glycans in the optimal spacing and orientation for target recognition, SELMA furnishes glycoaptamers with highly optimized glycan clustering, achieving low-nanomolar recognition. Although numerous applications can be envisioned, the protocols and discussions in this article describe procedures involved in applying SELMA to the discovery glycoDNAs that bind to the HIV broadly neutralizing antibody 2G12.