Rapid direct detection of the major fish allergen, parvalbumin, by selected MS/MS ion monitoring mass spectrometry

Abstract

Parvalbumins beta (β-PRVBs) are considered the major fish allergens. A new strategy for the rapid and direct detection of these allergens in any foodstuff is presented in this work.

An overview on fish and shellfish allergens and current methods of detection

“…The analysis of allergens by MS methodology can be performed by one of two approaches, either targeting intact proteins (analyte and reference standards) or peptides obtained from protein digestion using proteolytic enzymes (Picariello et al, 2011). This methodology was already applied for the direct detection of parvalbumin peptide biomarkers using a set of 16 species of fish that were analysed by LC-MS/MS approach (Carrera, Cañas, & Gallardo, 2012).…”

Detection of Allergen Markers in Food: Analytical Methods

“…Carrera et al [189,190] developed a rapid detection method for the purification of fi β-parvalbumin via heat treatment and accelerated in-solution trypsin digestion under an ultrasonic field. Peptide fi markers were monitored using selected ion monitoring MS and enabled the unequivocal identification of closely related fi fi sh species in processed fi seafood products.…”

High-Throughput Proteomics: A New Tool for Quality and Safety in Fishery Products

“…However, the authors were able to recognize clearly species-specific differences in the peptides pattern of the parvalbumin fractions only through MALDI-TOF mass fingerprinting and subsequently to identify 25 new ß-PRVBs isoforms by de novo MSsequencing in 11 species of Merluccidae through a classical bottom-up proteomic approach using Fourier-transform ioncyclotron resonance mass spectrometry (FTICR-MS) for an accurate mass measurement of intact proteins and selected MS/MS ion monitoring (SMIM) of peptide mass gaps [114]. Although these studies have been based on the preliminary use of 2DE, Carrera and colleagues [17] recently developed a new shotgun proteomic strategy more suitable for a rapid detection of fish ß-PRVBs in any food product in less than 2 hours and its validity in commercial sea-foodstuffs were tested in processed and even battered pre-cooked products. Briefly, this targeted-driven methodology is based on the fast purification of parvalbumins by heat treatment, accelerated in-solution protein digestion by high-intensity focused ultrasound (HIFU) monitoring of 19 ß-PRVBs common peptide biomarkers by SMIM in a linear ion trap (LIT) MS.…”

“…The major fish allergen (Parvalbumin beta, -PRVB), can now be detected in less than 2 hours using this technique [16,17]. This fast procedure can therefore be used to avoid fraudulent species substitution that can interfere with the quality and value of these products.…”