Xenosense Ltd., Belfast, Northern Ireland.
Talanta. 2011 May 15;84(3):638-43. doi: 10.1016/j.talanta.2011.01.036. Epub 2011 Jan 22.
A surface plasmon resonance (SPR) immunobiosensor assay was developed and validated to detect microcystin toxins in Spirulina and Aphanizomenon flos-aquae blue-green algae (BGA) food supplements. A competitive inhibition SPR-biosensor was developed using a monoclonal antibody to detect microcystin (MC) toxins. Powdered BGA samples were extracted with an aqueous methanolic solution, centrifuged and diluted in HBS-EP buffer prior to analysis. The assay was validated in accordance with the performance criteria outlined in EU legislation 2002/657/EC. The limit of detection (LOD) of the assay was calculated from the analysis of 20 known negative BGA samples to be 0.561 mg kg(-1). The detection capability (CCβ) of the assay was determined to be ≤ 0.85 mg kg(-1) for MC-LR. The biosensor assay was successfully applied to detect MC-LR toxins in BGA samples purchased on the Irish retail market. MC-LR was detected in samples at levels ranging from <0.5 to 2.21 mg kg(-1). The biosensor results were in good agreement with an established LC-MS/MS assay. The assay is advantageous because it employs a simple clean-up procedure compared to chemical assays and allows automated unattended analysis of samples unlike ELISA.
建立并验证了一种表面等离子体共振(SPR)免疫生物传感器分析方法,用于检测螺旋藻和鱼腥藻蓝绿藻(BGA)食品补充剂中的微囊藻毒素。使用单克隆抗体开发了竞争性抑制 SPR 生物传感器,以检测微囊藻毒素(MC)毒素。将粉末状 BGA 样品用含水甲醇溶液提取,离心,然后在 HBS-EP 缓冲液中稀释,再进行分析。该测定方法符合欧盟法规 2002/657/EC 中概述的性能标准进行了验证。通过对 20 个已知阴性 BGA 样品的分析,计算出该测定方法的检测限(LOD)为 0.561mg/kg。测定方法的检测能力(CCβ)确定为≤0.85mg/kg,用于 MC-LR。该生物传感器测定方法成功应用于检测爱尔兰零售市场上购买的 BGA 样品中的 MC-LR 毒素。在样品中检测到 MC-LR 的浓度范围为<0.5 至 2.21mg/kg。生物传感器的结果与建立的 LC-MS/MS 测定方法吻合良好。与化学测定方法相比,该测定方法采用了简单的净化程序,并且与 ELISA 不同,允许自动无人值守地分析样品,因此具有优势。