Nucleic Acid Ligands With Protein-like Side Chains: Modified Aptamers and Their Use as Diagnostic and Therapeutic Agents.
Nucleic Acid Ligands With Protein-like Side Chains: Modified Aptamers and Their Use as Diagnostic and Therapeutic Agents.
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DOI：
10.1038/mtna.2014.49
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发表时间：
2014-10-07
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核酸文库的有限化学多样性一直被怀疑是SELEX（指数富集配体系统进化）总体成功的主要限制因素。尽管有这种限制，但由于起始文库的巨大规模和核酸的构象丰富性，SELEX在过去的四分之一个世纪中取得了相当大的成功。通过明智地引入天然核酸中不存在的官能团，基于核酸的配体和基于蛋白质的配体之间的“多样性差距”可以基本上被桥接，以产生代表两个世界中最好的一类新的配体。我们已经探索了尿嘧啶5位上各种官能团的作用，发现疏水性芳族侧链对SELEX的成功率具有最深远的影响，并允许鉴定具有非常低的解离速率常数的配体（称为慢解离速率修饰的适体或SOMAmer）。这种修饰的核苷酸产生独特的分子内基序，并与蛋白质直接接触。重要的是，SOMAmer使其蛋白质靶点与比传统适体具有更大疏水性的表面结合，从而增加了可用于结合的表位范围。这些改进使我们能够建立超过3，000种人类蛋白质的SOMAmer集合，包括主要家族，如生长因子，细胞因子，酶，激素和受体，以及针对病原体和啮齿动物蛋白质的其他SOMAmer。如此大量且不断增长的精致亲和试剂的集合扩大了诊断和治疗中可能应用的范围。
Limited chemical diversity of nucleic acid libraries has long been suspected to be a major constraining factor in the overall success of SELEX (Systematic Evolution of Ligands by EXponential enrichment). Despite this constraint, SELEX has enjoyed considerable success over the past quarter of a century as a result of the enormous size of starting libraries and conformational richness of nucleic acids. With judicious introduction of functional groups absent in natural nucleic acids, the “diversity gap” between nucleic acid–based ligands and protein-based ligands can be substantially bridged, to generate a new class of ligands that represent the best of both worlds. We have explored the effect of various functional groups at the 5-position of uracil and found that hydrophobic aromatic side chains have the most profound influence on the success rate of SELEX and allow the identification of ligands with very low dissociation rate constants (named Slow Off-rate Modified Aptamers or SOMAmers). Such modified nucleotides create unique intramolecular motifs and make direct contacts with proteins. Importantly, SOMAmers engage their protein targets with surfaces that have significantly more hydrophobic character compared with conventional aptamers, thereby increasing the range of epitopes that are available for binding. These improvements have enabled us to build a collection of SOMAmers to over 3,000 human proteins encompassing major families such as growth factors, cytokines, enzymes, hormones, and receptors, with additional SOMAmers aimed at pathogen and rodent proteins. Such a large and growing collection of exquisite affinity reagents expands the scope of possible applications in diagnostics and therapeutics.
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DOI：
10.1007/978-1-59745-557-2_5
发表时间：
2009
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Cerchia, Laura;Giangrande, Paloma H;McNamara, James O;de Franciscis, Vittorio
通讯作者：
de Franciscis, Vittorio
影响因子：
14.9
作者：
AURUP, H;TUSCHL, T;ECKSTEIN, F
通讯作者：
ECKSTEIN, F
DOI：
10.1038/nsb0298-133
发表时间：
1998-02-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Convery, MA;Rowsell, S;Stockley, PG
通讯作者：
Stockley, PG
影响因子：
100.3
作者：
Beck, Alain;Wurch, Thierry;Corvaia, Nathalie
通讯作者：
Corvaia, Nathalie
影响因子：
--
作者：
Burmeister, PE;Lewis, SD;Keefe, AD
通讯作者：
Keefe, AD