Inter-laboratory optimization of protein extraction, separation, and fluorescent detection of endogenous rice allergens. Issue 11 (1st November 2016)

• Record Type: Journal Article
• Authors: Satoh, Rie; Teshima, Reiko; Kitta, Kazumi; Lang, Gang-Hua; Schegg, Kathleen; Blumenthal, Kenneth; Hicks, Leslie; Labory-Carcenac, Bénédicte; Rouquié, David; Herman, Rod A.; Herouet-Guicheney, Corinne; Ladics, Gregory S.; McClain, Scott; Poulsen, Lars K.; Privalle, Laura; Ward, Jason M.; Doerrer, Nancy; Rascle, Jean-Baptiste
• Is Part Of: Bioscience, biotechnology, and biochemistry. Volume 80:Issue 11(2016)
• Page Start: 2198
• Page End: 2207
• Publication Date: 2016-11-01
• Subjects: rice grain -- allergen -- proteomics -- multiplex immunodetection -- two-dimensional difference gel electrophoresis (2D-DIGE); Biotechnology -- Periodicals; Biochemistry -- Periodicals; 660.6
• DOI: 10.1080/09168451.2016.1206810
• Languages: English
• ISSNs: 0916-8451

Abstract

Abstract : 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. Graphical abstract: Representative multiplexed detection and 2D-DIGE profile of rice endogenous allergens.