Department of Analytical Chemistry, Faculty of Chemistry, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Anal Bioanal Chem. 2010 Nov;398(6):2625-34. doi: 10.1007/s00216-010-3856-8. Epub 2010 Jun 9.
A surface plasmon resonance (SPR) biosensor for the detection of microcystins (MCs) in drinking water has been developed. Several assay formats have been evaluated. The selected format is based on a competitive inhibition assay, in which microcystin-LR (MCLR) has been covalently immobilized onto the surface of an SPR chip functionalized with a self-assembled monolayer. The influence of several factors affecting sensor performance, such as the nature and concentration of the antibody, the composition of the carrier buffer, and the blocking and regeneration solutions, has been evaluated. The optimized SPR biosensor provides an IC(50) 0.67 ± 0.09 µg L(-1), a detection limit of 73 ± 8 ng L(-1), and a dynamic range from 0.2 to 2.0 µg L(-1) for MCLR. Cross-reactivity to other related MCs, such as microcystin-RR (88%) and microcystin-YR (94%), has also been measured. The SPR biosensor can perform four simultaneous determinations in 60 min, and each SPR chip can be reused for at least 40 assay-regeneration cycles without significant binding capacity loss. The biosensor has been successfully applied to the direct analysis of MCLR in drinking water samples, below the provisional guideline value of 1 µg L(-1) established by the World Health Organization for drinking water.
已开发出一种用于检测饮用水中微囊藻毒素 (MCs) 的表面等离子体共振 (SPR) 生物传感器。已评估了几种分析方法。选择的方法基于竞争抑制测定法，其中微囊藻毒素-LR (MCLR) 已通过自组装单层功能化的 SPR 芯片上的共价固定化。评估了影响传感器性能的几个因素，例如抗体的性质和浓度、载体缓冲液的组成以及封闭和再生溶液。优化后的 SPR 生物传感器为 MCLR 提供了 0.67 ± 0.09 µg L(-1) 的 IC(50)、73 ± 8 ng L(-1) 的检测限以及 0.2 至 2.0 µg L(-1) 的动态范围。还测量了对其他相关 MCs（如微囊藻毒素-RR（88%）和微囊藻毒素-YR（94%））的交叉反应性。SPR 生物传感器可以在 60 分钟内同时进行四次测定，并且每个 SPR 芯片在没有明显结合能力损失的情况下至少可以重复使用 40 次测定-再生循环。该生物传感器已成功应用于直接分析饮用水中的 MCLR，其含量低于世界卫生组织为饮用水设定的暂定指导值 1 µg L(-1)。
