MALDI-TOF MS and Magnetic Beads for Rapid Seafood Allergen Tests.

ABSTRACT

We developed a strategy using immunomagnetic separation (IMS) coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to test seafood allergens. The protocol employed commercial magnetic beads (MBs) functionalized with anti-human IgE antibodies to carry out the IMS of IgEs in blood samples, followed by capture of allergens from seafood protein extracts for allergy analysis. After elution, the captured allergens were identified by MALDI-TOF MS and HPLC-MS/MS. The non-specific adsorption of MBs to biomolecules, the reproducibility and sensitivity of the protocol were investigated. The method shows consistent results with enzyme-linked immunosorbent assay tests. The false positive rate of the present method for the allergy test is 0%. The protocol was applied to detect the allergens in greasy-back shrimp for checking the allergenicity of patients' serum. Cooking fish as soup may effectively decrease the allergenicity. The method can be potentially used to identify unknown allergens of seafood to ensure the safety of allergic patients.