de-los-Santos-Alvarez Noemí, Lobo-Castañón María Jesús, Miranda-Ordieres Arturo J, Tuñón-Blanco Paulino
Departamento de Química Física y Analítica, Universidad de Oviedo, Julián Clavería 8, 33006 Oviedo, Spain.
Biosens Bioelectron. 2009 Apr 15;24(8):2547-53. doi: 10.1016/j.bios.2009.01.011. Epub 2009 Jan 19.
We have studied how the modification of the RNA aptamer evolved against neomycin B at 2' position of ribose with a methyl group influences the affinity of the interaction. Using surface plasmon resonance (SPR) and faradaic impedance spectroscopy (FIS) an affinity constant in the muM range was calculated. The results showed that the modification of the aptamer does not significantly alter the affinity of the aptamer for the antibiotic. This finding opens up the possibility of designing modified RNA aptamers resistant to endonucleases without variation of the analytical features. In addition to this, we propose a competitive assay for the detection of neomycin B using SPR as a transduction technique. A range of quantification between 10 nM and 100 microM was obtained, which shows the feasibility of detecting small molecules using aptamers with high sensitivity.
我们研究了核糖2'位带有甲基的、针对新霉素B进化的RNA适配体修饰如何影响相互作用的亲和力。使用表面等离子体共振（SPR）和法拉第阻抗谱（FIS）计算出了微摩尔范围内的亲和常数。结果表明，适配体的修饰不会显著改变其对抗生素的亲和力。这一发现为设计对内切核酸酶具有抗性且不改变分析特性的修饰RNA适配体开辟了可能性。除此之外，我们提出了一种使用SPR作为转导技术检测新霉素B的竞争性分析方法。获得了10 nM至100 μM的定量范围，这表明使用适配体以高灵敏度检测小分子的可行性。
