Inter-laboratory optimization of protein extraction, separation, and fluorescent detection of endogenous rice allergens
Research output: Contribution to journal › Journal article › Research › peer-review
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 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 α-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.
Original language | English |
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Journal | Bioscience, Biotechnology, and Biochemistry |
Volume | 80 |
Issue number | 11 |
Pages (from-to) | 2198-2207 |
Number of pages | 10 |
ISSN | 0916-8451 |
DOIs | |
Publication status | Published - 2016 |
ID: 165662413