Department of Chemistry, Masaryk University, Kotlarska 2, CZ-611 37 Brno, Czech Republic.
Biosens Bioelectron. 2011 Jan 15;26(5):2201-7. doi: 10.1016/j.bios.2010.09.035.
In this paper, heavy metal biosensor based on immobilization of metallothionein (MT) to the surface of carbon paste electrode (CPE) via anti-MT-antibodies is reported. First, the evaluation of MT electroactivity was done. The attention was focused on the capturing of MT to the CPE surface. Antibodies incorporated and mixed into carbon paste were stable; even after two weeks the observed changes in signal height were lower than 5%. Further, the interaction of MT with polyclonal chicken antibodies incorporated in carbon paste electrode was determined by square-wave voltammetry. In the voltammogram, two signals--labelled as cys(MT) and W(a)--were observed. The cys(MT) corresponded to -SH moieties of MT and W(a) corresponded to tryptophan residues of chicken antibodies. Time of interaction (300 s) and MT concentration (125 μg/ml) were optimized to suggest a silver(I) ions biosensor. Biosensor (CPE modified with anti-MT antibody) prepared under the optimized conditions was then used for silver(I) ions detection. The detection limit (3 S/N) for silver(I) ions was estimated as 0.5 nM. The proposed biosensor was tested by detection spiking of silver(I) ions in various water samples (from very pure distilled water to rainwater). Recoveries varied from 74 to 104%.
本文报道了一种基于重金属生物传感器,通过抗 MT 抗体将金属硫蛋白(MT)固定在碳糊电极(CPE)表面的方法。首先,评估了 MT 的电化学活性。研究重点是 MT 被捕获到 CPE 表面。掺入和混合到碳糊中的抗体是稳定的;即使在两周后,观察到的信号高度变化也低于 5%。此外,通过方波伏安法测定了 MT 与掺入碳糊电极中的多克隆鸡抗体的相互作用。在伏安图中,观察到两个信号--标记为 cys(MT)和 W(a)。cys(MT)对应于 MT 的 -SH 部分,W(a)对应于鸡抗体的色氨酸残基。优化了相互作用时间(300 s)和 MT 浓度(125 μg/ml),以提出一种银(I)离子生物传感器。在优化条件下制备的生物传感器(用抗 MT 抗体修饰的 CPE)随后用于银(I)离子的检测。银(I)离子的检测限(3 S/N)估计为 0.5 nM。该生物传感器通过在各种水样(从非常纯净的蒸馏水到雨水)中检测银(I)离子的加标来测试。回收率从 74%到 104%不等。