Analysis of the potential allergenicity of traditional apple cultivars by Multiplex Biochip-Based Immunoassay

Potential allergenicity of Medicago sativa investigated by a combined IgE‐binding inhibition, proteomics and in silico approach

“…Protein extract was prepared from the dried powder of alfalfa using the protein extraction procedure developed for plant foods 11 with a few modifications. Briefly, 20 g of ground dried leaves were homogenized in a solution made with 80 mL of 0.5 mol L ‐1 sodium chloride (NaCl), 0.8 g of poly(vinylpolypyrrolidone), 2 mL of 0.125 mol L ‐1 ethylenediaminetetraacetic acid and 2.2 g of ascorbic acid dissolved in 25 mL of water and brought to pH 3.5 by addition of sodium hydroxide (NaOH).…”

“…For instance, it can be used to investigate immunological similarities between different molecules and also to detect possible allergens in a protein mixture, such as a food extract, by performing IgE inhibition test. In food allergen testing, multiplex systems that allow the analysis of a comprehensive panel of possible allergens in the proteome of a food extract are especially of value 11–13 . Pasquariello et al ., for example, have used IgE inhibition tests performed with the Single Point Highest Inhibition Achievable assay (SPHIAa) in combination with a multiplex biochip‐based immunoassay to analyse the allergen profiles of protein extracts from different apple cultivars 11 and to select potentially hypo‐ or hyper‐allergenic apple cultivars.…”

“…In food allergen testing, multiplex systems that allow the analysis of a comprehensive panel of possible allergens in the proteome of a food extract are especially of value 11–13 . Pasquariello et al ., for example, have used IgE inhibition tests performed with the Single Point Highest Inhibition Achievable assay (SPHIAa) in combination with a multiplex biochip‐based immunoassay to analyse the allergen profiles of protein extracts from different apple cultivars 11 and to select potentially hypo‐ or hyper‐allergenic apple cultivars. Similarly, the IgE inhibition with the SPHIAa method, combined with the allergen multiplex FABER test, was used to assess the cross‐reaction between a protein extract and individual allergens, namely the cypress pollen extract and the peach and cypress GRPs 12 …”

“…1) combines proteomics‐based protein identification, immunological investigations, bioinformatics 30 and data mining to gain biological insights useful to evaluate the presence of potential allergens in protein extract. The possible allergenicity of the leaf protein extract of M. sativa L was investigated by IgE‐binding inhibition test using the SPHIAa method 11,12,31,32 associated to FABER test 12 . FABER is a nanotechnology‐based multiplex in vitro serological test, which takes advantage of database and bioinformatics tools of the Allergome platform (http://www.allergome.org/) to analyze and store diagnostic and research data.…”

Plant-Made Bet v 1 for Molecular Diagnosis

“…More importantly, it frequently fails to detect specific IgE because the composition of the surrogate markers can be very variable (Giangrieco et al, 2012). This high variability makes it impossible to have standardized extracts with a constant allergen composition, since this may be influenced by several factors, such as differences among plant cultivars, post-harvest treatments, specific phenological stages, and extraction protocols (Ciardiello et al, 2009; Pasquariello et al, 2012). Molecule-based diagnosis may overcome all these drawbacks, given the possibility of detecting the specific IgE against individual allergenic molecules (Giangrieco et al, 2012; Ciardiello et al, 2013).…”