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Biosens Bioelectron. 2013 Dec 15;50:150-6. doi: 10.1016/j.bios.2013.06.032. Epub 2013 Jun 24.

Mast cell-based electrochemical biosensor for quantification of the major shrimp allergen Pen a 1 (tropomyosin).

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  • 1State Key Laboratory of Food Science and Technology, School of Food Science of Jiangnan University, Wuxi, Jiangsu 214122, China.

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.

Copyright © 2013 Elsevier B.V. All rights reserved.

KEYWORDS:

Collagen; Electrochemical sensor; RBL-2H3 mast cells; Shrimp tropomyosin

PMID:
23850781
[PubMed - indexed for MEDLINE]
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