title: Fluorescent magnetic bead-based mast cell biosensor for electrochemical detection of allergens in foodstuffs.
abstract: 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.
authors.affiliation: 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.
authors.forename: Donglei
authors.initials: D
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Dairy Biotechnology, Dairy Research Institute, Bright Dairy & Food Co., Ltd., Shanghai 200436, China.
authors.forename: Pei
authors.initials: P
authors.lastname: Zhu
authors.affiliation: 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.
authors.forename: Hui
authors.initials: H
authors.lastname: Jiang
authors.affiliation: 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.
authors.forename: Jian
authors.initials: J
authors.lastname: Ji
authors.affiliation: 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. Electronic address: sxlzzz@jiangnan.edu.cn.
authors.forename: Xiulan
authors.initials: X
authors.lastname: Sun
authors.affiliation: 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.
authors.forename: Wenshu
authors.initials: W
authors.lastname: Gu
authors.affiliation: 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.
authors.forename: Genyi
authors.initials: G
authors.lastname: Zhang
mesh_terms: D000485:Allergens / Q000032:analysis* / Q000276:immunology; D000818:Animals; D001681:Biological Assay / Q000295:instrumentation; D015374:Biosensing Techniques / Q000295:instrumentation*; D002460:Cell Line; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005504:Food Analysis / Q000295:instrumentation*; D005506:Food Contamination / Q000032:analysis*; D018189:Immunomagnetic Separation / Q000295:instrumentation*; D008407:Mast Cells / Q000187:drug effects* / Q000276:immunology; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000295:instrumentation
chemical_list: D000485:Allergens
keywords: Electrochemical impedance spectroscopy; FITC; Fe(3)O(4); Fish parvalbumin; Lipidosome; RBL-2H3 mast cells; Shrimp tropomyosin Pen a 1; SiO(2)