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

ABSTRACT: In rice, several allergens have been identified such as the non-specific lipid transfer protein-1, the ?-amylase/trypsin-inhibitors, the ?-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 ring-trial experiment, several protein extraction methods were first compared and analyzed by 1D multiplexed SDS-PAGE. In a second phase, an inter-laboratory 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 ?-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.

SUBMITTER: Satoh R

PROVIDER: S-EPMC5062055 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature

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

Satoh Rie R Teshima Reiko R Kitta Kazumi K Lang Gang-Hua GH Schegg Kathleen K Blumenthal Kenneth K Hicks Leslie L Labory-Carcenac Bénédicte B Rouquié David D Herman Rod A RA Herouet-Guicheney Corinne C Ladics Gregory S GS McClain Scott S Poulsen Lars K LK Privalle Laura L Ward Jason M JM Doerrer Nancy N Rascle Jean-Baptiste JB

Bioscience, biotechnology, and biochemistry 20160711 11

In rice, several allergens have been identified such as the non-specific lipid transfer protein-1, the α-amylase/trypsin-inhibitors, the α-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 ring-trial experiment, several protein extr ...[more]

PMID: 27399872

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Dataset Information

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

Publications

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

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