School of Food Science and Technology, State Key Laboratory of Food Science and Technology, Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, Jiangsu 214122, China.
State Key Laboratory of Dairy Biotechnology, Dairy Research Institute, Bright Dairy & Food Co., Ltd., Shanghai 200436, China.
Biosens Bioelectron. 2015 Aug 15;70:482-90. doi: 10.1016/j.bios.2015.03.058. Epub 2015 Mar 28.
In this study, a novel electrochemical rat basophilic leukemia cell (RBL-2H3) cell sensor, based on fluorescent magnetic beads, has been developed for the detection and evaluation of different allergens in foodstuffs. Fluorescein isothiocyanate (FITC) was successfully fused inside the SiO2 layer of SiO2 shell-coated Fe3O4 nanoparticles, which was superior to the traditional Fe3O4@SiO2@FITC modification process. The as-synthesized fluorescent magnetic beads were then encapsulated with lipidosome to form cationic magnetic fluorescent nanoparticles (CMFNPs) for mast cell magnetofection. The CMFNPs were then characterized by SEM, TEM, VSM, FTIR, and XRD analyses, and transfected into RBL-2H3 cells through a highly efficient, lipid-mediated magnetofection procedure. Magnetic glassy carbon electrode (MGCE), which possesses excellent reproducibility and regeneration qualities, was then employed to adsorb the CMFNP-transfected RBL-2H3 cells activated by an allergen antigen for electrochemical assay. Results show that the exposure of model antigen-dinitrophenol-bovine serum albumin (DNP-BSA) to anti-DNP IgE-sensitized mast cells induced a robust and long-lasting electrochemical impedance signal in a dose-dependent manner. The detection limit was identified at 3.3×10(-4) ng/mL. To demonstrate the utility of this mast cell-based biosensor for detection of real allergens in foodstuffs, Anti-Pen a1 IgE and Anti-PV IgE-activated cells were employed to quantify both shrimp allergen tropomyosin (Pen a 1) and fish allergen parvalbumin (PV). Results show high detection accuracy for these targets, with a limit of 0.03 μg/mL (shrimp Pen a 1) and 0.16 ng/mL (fish PV), respectively. To this effect, we conclude the proposed method is a facile, highly sensitive, innovative electrochemical method for the evaluation of food allergens.
在这项研究中，基于荧光磁性珠，开发了一种新型电化学大鼠嗜碱性白血病细胞 (RBL-2H3) 细胞传感器，用于检测和评估食品中的不同过敏原。异硫氰酸荧光素 (FITC) 成功地融合在 SiO2 壳层包裹的 Fe3O4 纳米粒子的 SiO2 层内，优于传统的 Fe3O4@SiO2@FITC 修饰过程。然后，将合成的荧光磁性珠用脂质体包封形成阳离子磁性荧光纳米颗粒 (CMFNPs) 用于肥大细胞磁转染。然后通过 SEM、TEM、VSM、FTIR 和 XRD 分析对 CMFNPs 进行表征，并通过高效的脂质介导的磁转染程序转染到 RBL-2H3 细胞中。然后，将具有优异重现性和再生质量的磁性玻碳电极 (MGCE) 用于吸附过敏原抗原激活的 CMFNP 转染的 RBL-2H3 细胞进行电化学测定。结果表明，模型抗原二硝基苯酚-牛血清白蛋白 (DNP-BSA) 暴露于抗 DNP IgE 敏化的肥大细胞中，以剂量依赖的方式诱导出强大而持久的电化学阻抗信号。检测限确定为 3.3×10(-4) ng/mL。为了证明基于肥大细胞的生物传感器用于检测食品中真实过敏原的实用性，使用 Anti-Pen a1 IgE 和 Anti-PV IgE 激活的细胞来定量虾过敏原原肌球蛋白 (Pen a 1) 和鱼过敏原副肌球蛋白 (PV)。结果表明，对这些靶标的检测具有很高的准确性，虾过敏原原肌球蛋白的检测限为 0.03 μg/mL，鱼过敏原副肌球蛋白的检测限为 0.16 ng/mL。因此，我们得出结论，该方法是一种简便、高灵敏度、创新的电化学方法，用于评估食物过敏原。
