Edogun Osafanmwen, Chan Tracy Y, Nguyen Nghia H, Luu Anthony, Halim Marlin
Department of Chemistry and Biochemistry, California State University, East Bay.
Department of Chemistry and Biochemistry, California State University, East Bay;
J Vis Exp. 2017 Jan 9(119):55216. doi: 10.3791/55216.
A method for determining the presence of unchelated trivalent gadolinium ion (Gd) in aqueous solution is demonstrated. Gd is often present in samples of gadolinium-based contrast agents as a result of incomplete reactions between the ligand and the ion, or as a dissociation product. Since the ion is toxic, its detection is of critical importance. Herein, the design and usage of an aptamer-based sensor (Gd-sensor) for Gd are described. The sensor produces a fluorescence change in response to increasing concentrations of the ion, and has a limit of detection in the nanomolar range (100 nM with a signal-to-noise ratio of 3). The assay may be run in an aqueous buffer at ambient pH (7 - 7.4) in a 384-well microplate. The sensor is relatively unreactive toward other physiologically relevant metal ions such as sodium, potassium, and calcium ions, although it is not specific for Gd over other trivalent lanthanides such as europium(III) and terbium(III). Nevertheless, the lanthanides are not commonly found in contrast agents or the biological systems, and the sensor may therefore be used to selectively determine unchelated Gd in aqueous conditions.
本文展示了一种测定水溶液中未螯合三价钆离子(Gd)的方法。由于配体与离子之间反应不完全,或作为解离产物,Gd通常存在于钆基造影剂样品中。由于该离子有毒,其检测至关重要。在此,描述了一种基于适配体的Gd传感器(Gd传感器)的设计和用途。该传感器会随着离子浓度的增加产生荧光变化,检测限在纳摩尔范围内(约100 nM,信噪比为3)。该测定可在384孔微孔板中,于环境pH值(约7 - 7.4)的水性缓冲液中进行。该传感器对其他生理相关金属离子(如钠、钾和钙离子)相对无反应,尽管它对Gd的特异性并不高于其他三价镧系元素(如铕(III)和铽(III))。然而,镧系元素在造影剂或生物系统中并不常见,因此该传感器可用于在水性条件下选择性测定未螯合的Gd。