Department of Chemistry, Tsinghua University, Beijing, China.
Anal Chem. 2012 Mar 20;84(6):2916-22. doi: 10.1021/ac203488p. Epub 2012 Mar 7.
In this work, two methods with complementary features, catalytic and molecular beacon (CAMB) and label-free fluorescent sensors using an abasic site, have been combined into new label-free CAMB sensors that possess advantages of each method. The label-free method using a dSpacer-containing molecular beacon makes CAMB more cost-effective and less interfering with the catalytic activity, while CAMB allows the label-free method to use true catalytic turnovers for signal amplifications, resulting in a new label-free CAMB sensor for Pb(2+) ion, with a detection limit of 3.8 nM while maintaining the same selectivity. Furthermore, by using CAMB to overcome the label-free method's limitation of requiring excess enzyme strands, a new label-free CAMB sensor using aptazyme is also designed to detect adenosine down to 1.4 μM, with excellent selectivity over other nucleosides.
在这项工作中,两种具有互补特征的方法——催化和分子信标 (CAMB) 以及基于无标记荧光传感器的碱基缺失位点,被结合到新的无标记 CAMB 传感器中,这些传感器具有每种方法的优点。使用含 dSpacer 的分子信标进行无标记方法,使 CAMB 更具成本效益,并且对催化活性的干扰更小,而 CAMB 允许无标记方法使用真正的催化循环进行信号放大,从而产生用于检测 Pb(2+)离子的新的无标记 CAMB 传感器,检测限为 3.8 nM,同时保持相同的选择性。此外,通过使用 CAMB 克服无标记方法需要过量酶链的限制,还设计了一种使用适体酶的新的无标记 CAMB 传感器,可检测低至 1.4 μM 的腺苷,对其他核苷具有出色的选择性。