Cranfield Health, Cranfield University, Cranfield, Bedfordshire, MK43 0AL, England, United Kingdom.
Environ Sci Technol. 2012 May 15;46(10):5504-10. doi: 10.1021/es2041042. Epub 2012 Apr 24.
We describe within this paper the development of an affinity sensor for the detection of the cyanobacterial toxin microcystin-LR. The first stage of the work included acquiring and testing of the antibodies to this target. Following the investigation, a heterogeneous direct competitive enzyme-linked immunosorbent assay (ELISA) format for microcystin-LR detection was developed, achieving a detection limit, LLD(80) = 0.022 μg L(-1). The system was then transferred to an affinity membrane sorbent-based ELISA. This was an amenable format for immunoassay incorporation into a disposable amperometric immunosensor device. This membrane-based ELISA achieved a detection limit, LLD(80) = 0.06 μg L(-1). A three-electrode immunosensor system was fabricated using thick-film screen-printing technology. Amperometric horseradish peroxidase transduction of hydrogen peroxide catalysis, at low reducing potentials, versus Ag/AgCl reference and carbon counter electrodes, was facilitated by hydroquinone-mediated electron transfer. A detection limit of 0.5 μg L(-1) for microcystin-LR was achieved. Similar levels of detection could be obtained using direct electrochemical sensing of the dye produced using the membrane-based ELISA. These techniques proved to be simple, cost-effective, and suitable for the detection of microcystin-LR in buffer and spiked tap and river water samples.
本文描述了一种用于检测蓝藻毒素微囊藻毒素-LR 的亲和传感器的开发。这项工作的第一阶段包括获取和测试针对该目标的抗体。经过调查，开发了一种用于微囊藻毒素-LR 检测的异质直接竞争酶联免疫吸附测定（ELISA）格式，检测限 LLD(80) = 0.022 μg L(-1)。然后，该系统被转移到基于亲和膜吸附剂的 ELISA 中。这是一种适合将免疫测定纳入一次性电流免疫传感器设备的格式。这种基于膜的 ELISA 实现了检测限 LLD(80) = 0.06 μg L(-1)。使用厚膜丝网印刷技术制造了三电极免疫传感器系统。通过氢醌介导的电子转移，在低还原电势下，辣根过氧化物酶对过氧化氢的催化进行了电流测定，相对于 Ag/AgCl 参比电极和碳对电极。微囊藻毒素-LR 的检测限为 0.5 μg L(-1)。使用基于膜的 ELISA 直接电化学检测生成的染料也可以获得类似的检测水平。这些技术被证明简单、具有成本效益，并且适合于缓冲液和加标自来水和河水样品中微囊藻毒素-LR 的检测。
