Investigation on Sequential Extraction of Peanut Allergens for Subsequent Analysis by ELISA and 2D Gel Electrophoresis
A two-step sequential extraction method of peanut proteins was proposed with the aim to investigate the protein composition and
allergen content of peanut samples. The extraction procedure reported is fully compatible with subsequent analysis by enzyme-linked
immunosorbent assays (ELISA) as well as 2D gel electrophoresis (2D PAGE). This sequential extraction method was used to study three
different peanut varieties and three different types of food processing. Peanuts were analysed for total protein content and the extraction
efficiency of raw and processed peanuts was determined. The total protein content of the three peanut varieties was found to be comparable,
but their extraction efficiency varies. The peanut extracts were characterised by employing three different ELISA test kits specific
to either the allergens Ara h 1 or Ara h 2, or to soluble peanut proteins. The content of both Ara h 1 and Ara h 2 differed in the raw
peanut extracts of the three varieties. However, thermal processing resulted in much larger changes in detectability. Blanching significantly
increases the detectability of Ara h 2, whereas Ara h 1 detection remains almost unchanged. After roasting a clear decrease of
detectability was observed for both Ara h 1 and Ara h 2, although the effect is more severe for Ara h 1. 2D PAGE was employed to
compare the protein profiles and abundances of peanut extracts. Statistically relevant differences were observed for the two different protein
fractions obtained by using the described method, showing the relevance of this two-step sequential extraction method.
CHASSAIGNE Hubert;
BROHEE Marcel;
NORGAARD Jorgen;
VAN HENGEL Adrianus;
2007-10-08
ELSEVIER SCI LTD
JRC40113
0308-8146,
https://publications.jrc.ec.europa.eu/repository/handle/JRC40113,
10.1016/j.foodchem.2007.04.055,
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