title: Mast cell-based electrochemical biosensor for quantification of the major shrimp allergen Pen a 1 (tropomyosin).
abstract: A novel cell-based electrochemical biosensor was developed to quantify major shrimp allergen Pen a 1 (tropomyosin) and to assess its immunoglobulin E (IgE)-mediated hypersensitivity. Rat basophilic leukemia (RBL-2H3) mast cells, encapsulated in type I collagen, were immobilized on a self-assembled l-cysteine/gold nanoparticle (AuNPsCys)-modified gold electrode to monitor IgE-mediated mast cell sensitization and activation. The exposure of dinitrophenol-bovine serum albumin (DNP-BSA), as a model antigen that stimulates mast cells, induced a robust and long-lasting electrochemical impedance signal in a dose-dependent manner which efficiently measured degranulation of anti-DNP IgE-stimulated mast cells. Then this mast cell-based biosensor was applied into quantification for the shrimp allergen with anti-shrimp tropomyosin IgE-sensitization. The electrochemical impedance spectroscopy (EIS) results showed that the impedance value (Ret) increased with the concentration of purified shrimp allergen Pen a 1 (tropomyosin) in range of 0.5-0.25 μg mL(-1) with the detection limit as 0.15 μg mL(-1), and the electrochemical result was confirmed by β-hexosaminidase assay and scanning electron microscopic morphological (SEM) analysis. Thus, a simple, label-free, and sensitive method for the determination of shrimp allergens was proposed and demonstrated here, implying a highly versatile biosensor for food allergen detection and prediction.
authors.affiliation: State Key Laboratory of Food Science and Technology, School of Food Science of Jiangnan University, Wuxi, Jiangsu 214122, China.
authors.forename: Donglei
authors.initials: D
authors.lastname: Jiang
authors.forename: Jian
authors.initials: J
authors.lastname: Ji
authors.forename: Lu
authors.initials: L
authors.lastname: An
authors.forename: Xiulan
authors.initials: X
authors.lastname: Sun
authors.forename: Yinzhi
authors.initials: Y
authors.lastname: Zhang
authors.forename: Genyi
authors.initials: G
authors.lastname: Zhang
authors.forename: Lili
authors.initials: L
authors.lastname: Tang
mesh_terms: D000485:Allergens / Q000032:analysis* / Q000276:immunology; D000818:Animals; D060829:Arthropod Proteins / Q000032:analysis* / Q000276:immunology; D015374:Biosensing Techniques / Q000295:instrumentation*; D002460:Cell Line; D055664:Electrochemical Techniques / Q000295:instrumentation*; D004867:Equipment Design; D005504:Food Analysis / Q000295:instrumentation*; D057230:Limit of Detection; D008407:Mast Cells / Q000187:drug effects / Q000276:immunology; D033561:Penaeidae / Q000737:chemistry* / Q000276:immunology; D051381:Rats; D014335:Tropomyosin / Q000032:analysis*
chemical_list: D000485:Allergens; D060829:Arthropod Proteins; D014335:Tropomyosin; C058449:shrimp allergens Sa
keywords: Collagen; Electrochemical sensor; RBL-2H3 mast cells; Shrimp tropomyosin