Tilapia (Oreochromis mossambicus) allergens characterized by ELISA, SDS-PAGE, 2D gels, Western blotting and MALDI-TOF mass spectrometry

Abstract

Parvalbumins have long been identified as major allergens in fish, but our research has found that parvalbumins are not the main cause of allergic reactions to tilapia. After homogenization, proteins were extracted from freshwater tilapia to react with a pooled sera sample taken from patients (n = 20) tested to be allergic to tilapia. Enzyme-linked immunosorbent assay revealed immunoglobulin E antibody activity, followed by sodium dodecyl sulphate polyacrylamide gel electrophoresis, Western blot, 2D electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to carry out protein separation and analysis. Protein identification against different databases yielded three known high molecular weight proteins as tilapia allergens: chromosome undetermined SCAF7145, fructose-bisphosphate aldolase A and enolase 3 (beta muscle). A fourth, new, unidentified protein with two T-cell receptors was discovered.

Keywords::

• fish
• 2D SDS-PAGE
• MALDI-TOF
• ELISA

Declaration of interest: The authors report no conflict of interest. The authors alone are responsible for the content and writing of the paper.