Inter-laboratory optimization of protein extraction, separation, and fluorescent detection of endogenous rice allergens

Rie Satoh, Reiko Teshima, Kazumi Kitta, Gang-Hua Lang, Kathleen Schegg, Kenneth Blumenthal, Leslie Hicks, Bénédicte Labory-Carcenac, David Rouquié, Rod A Herman, Corinne Herouet-Guicheney, Gregory S Ladics, Scott McClain, Lars K Poulsen, Laura Privalle, Jason M Ward, Nancy Doerrer, Jean-Baptiste Rascle

3 Citationer (Scopus)

Abstract

In rice, several allergens have been identified such as the non-specific lipid transfer protein-1, the α-amylase/trypsin-inhibitors, the á-globulin, the 33 kDa glyoxalase I (Gly I), the 52-63 kDa globulin, and the granule-bound starch synthetase. The goal of the present study was to define optimal rice extraction and detection methods that would allow a sensitive and reproducible measure of several classes of known rice allergens. In a three-laboratory ringtrial experiment, several protein extraction methods were first compared and analyzed by 1D multiplexed SDS-PAGE. In a second phase, an interlaboratory validation of 2D-DIGE analysis was conducted in five independent laboratories, focusing on three rice allergens (52 kDa globulin, 33 kDa glyoxalase I, and 14-16 kDa á-amylase/trypsin inhibitor family members). The results of the present study indicate that a combination of 1D multiplexed SDS-PAGE and 2D-DIGE methods would be recommended to quantify the various rice allergens.

OriginalsprogEngelsk
TidsskriftBioscience, Biotechnology, and Biochemistry
Vol/bind80
Udgave nummer11
Sider (fra-til)2198-2207
Antal sider10
ISSN0916-8451
DOI
StatusUdgivet - 2016

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