A DNA aptamer with high affinity and specificity for therapeutic anthracyclines
A DNA aptamer with high affinity and specificity for therapeutic anthracyclines
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DOI：
10.1016/j.ab.2007.09.007
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发表时间：
2008-02-01
影响因子：
2.9
通讯作者：
Gloekler, Joern
中科院分区：
文献类型：
--
作者：
Wochner, Aniela;Menger, Marcus;Gloekler, Joern
我们描述了表征的DNA适体，显示高亲和力和特异性的蒽环类药物柔红霉素和阿霉素，这两者都是经常用于化疗。使用用于磁珠的半自动化程序从随机序列库中分离适体。所有选择的适体显示出对靶分子柔红霉素的高亲和力。一种适体被进一步表征，并表现出20 nM的解离常数（K-D）。为了检查适体的结合特性并阐明其用于诊断测定的适用性，评价了其在各种缓冲液条件下的性能。适体被证明是非常稳健的，不依赖于特定离子的存在。它还耐受宽的pH范围和通过5 '-生物素化的固定。建立了检测柔红霉素的竞争法。这不仅允许测定适体的特异性，而且允许定量低至8.4 μ g/L的道诺霉素和阿霉素。(c)2007年爱思唯尔公司All rights reserved.
We describe the characterization of a DNA aptamer that displays high affinity and specificity for the anthracyclines daunomycin and doxorubicin, both of which are frequently used in chemotherapy. Aptamers were isolated from a pool of random sequences using a semi-automated procedure for magnetic beads. All selected aptamers displayed high affinity for the target molecule daunomycin. One aptamer was further characterized and exhibited a dissociation constant (K-D) of 20 nM. To examine the aptamer's binding properties and clarify its applicability for diagnostic assays, its performance under various buffer conditions was evaluated. The aptamer proved to be very robust and not dependent on the presence of specific ions. It also tolerated a wide pH range and immobilization via 5'-biotinylation. Furthermore, a competition assay for sensitive daunomycin detection was established. This not only allows the determination of the aptamer's specificity but also allows the quantification of as little as 8.4 mu g/L daunomycin and doxorubicin. (c) 2007 Elsevier Inc. All rights reserved.