TY - JOUR
T1 - Inter-laboratory optimization of protein extraction, separation, and fluorescent detection of endogenous rice allergens
AU - Satoh, Rie
AU - Teshima, Reiko
AU - Kitta, Kazumi
AU - Lang, Gang-Hua
AU - Schegg, Kathleen
AU - Blumenthal, Kenneth
AU - Hicks, Leslie
AU - Labory-Carcenac, Bénédicte
AU - Rouquié, David
AU - Herman, Rod A
AU - Herouet-Guicheney, Corinne
AU - Ladics, Gregory S
AU - McClain, Scott
AU - Poulsen, Lars K
AU - Privalle, Laura
AU - Ward, Jason M
AU - Doerrer, Nancy
AU - Rascle, Jean-Baptiste
PY - 2016
Y1 - 2016
N2 - 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.
AB - 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.
U2 - 10.1080/09168451.2016.1206810
DO - 10.1080/09168451.2016.1206810
M3 - Journal article
C2 - 27399872
SN - 0916-8451
VL - 80
SP - 2198
EP - 2207
JO - Bioscience, Biotechnology and Biochemistry
JF - Bioscience, Biotechnology and Biochemistry
IS - 11
ER -