2 research outputs found
Inter-laboratory optimization of protein extraction, separation, and fluorescent detection of endogenous rice allergens
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 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 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