Cranfield University, Advanced Diagnostics and Sensors Group, Cranfield, Bedford, MK43 0AL UK.
University of Manchester, School of Chemical Engineering and Analytical Science, Oxford Road, Manchester, M13 9PL UK.
Biosens Bioelectron. 2017 Feb 15;88:109-113. doi: 10.1016/j.bios.2016.07.101. Epub 2016 Jul 28.
A simple, sensitive and label-free optical sensor method was developed for allergens analysis using α-casein as the biomarker for cow's milk detection, to be used directly in final rinse samples of cleaning in place systems (CIP) of food manufacturers. A Surface Plasmon Resonance (SPR) sensor chip consisting of four sensing arrays enabling the measurement of samples and control binding events simultaneously on the sensor surface was employed in this work. SPR offers several advantages in terms of label free detection, real time measurements and superior sensitivity when compared to ELISA based techniques. The gold sensor chip was used to immobilise α-casein-polyclonal antibody using EDC/NHS coupling procedure. The performance of the assay and the sensor was first optimised and characterised in pure buffer conditions giving a detection limit of 58ngmL as a direct binding assay. The assay sensitivity can be further improved by using sandwich assay format and amplified with nanoparticles. However, at this stage this is not required as the detection limit achieved exceeded the required allergens detection levels of 2µgmL for α-S1-casein. The sensor demonstrated good selectivity towards the α-casein as the target analyte and adequate recoveries from CIP final rinse wash samples. The sensor would be useful tool for monitoring allergen levels after cleaning procedures, providing additional data that may better inform upon wider food allergen risk management decision(s) that are made by food manufacturer. In particular, this sensor could potentially help validate or optimise cleaning practices for a given food manufacturing process.
本研究开发了一种简单、灵敏且无需标记的光学传感器方法,用于分析过敏原,以α-酪蛋白作为牛乳检测的生物标志物,可直接用于食品制造商原位清洗(CIP)系统的最终冲洗样品中。本工作采用了一种表面等离子体共振(SPR)传感器芯片,该芯片由四个传感阵列组成,可同时在传感器表面上测量样品和控制结合事件。与基于 ELISA 的技术相比,SPR 在无标记检测、实时测量和更高的灵敏度方面具有多项优势。金传感器芯片通过 EDC/NHS 偶联程序用于固定化α-酪蛋白多克隆抗体。首先在纯缓冲条件下优化和表征了该测定法和传感器,作为直接结合测定法,检测限为 58ngmL。通过使用三明治测定法并进行纳米粒子放大,可以进一步提高测定的灵敏度。然而,在现阶段,由于检测限达到了α-S1-酪蛋白 2μgmL 的过敏原检测要求,因此不需要进一步提高灵敏度。该传感器对目标分析物α-酪蛋白具有良好的选择性,并且从 CIP 最终冲洗清洗样品中获得了足够的回收率。该传感器将成为监测清洁程序后过敏原水平的有用工具,提供可能更好地告知食品制造商做出更广泛的食物过敏原风险管理决策的附加数据。特别是,该传感器可以帮助验证或优化特定食品制造过程的清洁实践。