title: 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 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.
authors.affiliation: a Food Research Institute, National Agriculture and Food Research Organization , Tsukuba , Japan.
authors.forename: Rie
authors.initials: R
authors.lastname: Satoh
authors.affiliation: b Division of Novel Foods and Immunochemistry , National Institute of Health Sciences , Tokyo , Japan.
authors.forename: Reiko
authors.initials: R
authors.lastname: Teshima
authors.affiliation: a Food Research Institute, National Agriculture and Food Research Organization , Tsukuba , Japan.
authors.forename: Kazumi
authors.initials: K
authors.lastname: Kitta
authors.affiliation: a Food Research Institute, National Agriculture and Food Research Organization , Tsukuba , Japan.
authors.forename: Gang-Hua
authors.initials: GH
authors.lastname: Lang
authors.affiliation: c Nevada Proteomics Center , University of Nevada Reno , Reno , NV , USA.
authors.forename: Kathleen
authors.initials: K
authors.lastname: Schegg
authors.affiliation: d Formerly at Department of Biochemistry , State University of New York at Buffalo , Buffalo , NY , USA.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Blumenthal
authors.affiliation: e Formerly at Proteomics & Mass Spectrometry Facility , Donald Danforth Plant Science Center , St. Louis , MI , USA.
authors.forename: Leslie
authors.initials: L
authors.lastname: Hicks
authors.affiliation: f Bayer S.A.S., Bayer CropScience , Sophia Antipolis , France.
authors.forename: Bénédicte
authors.initials: B
authors.lastname: Labory-Carcenac
authors.affiliation: f Bayer S.A.S., Bayer CropScience , Sophia Antipolis , France.
authors.forename: David
authors.initials: D
authors.lastname: Rouquié
authors.affiliation: g Dow AgroSciences LLC , Indianapolis , IN , USA.
authors.forename: Rod A
authors.initials: RA
authors.lastname: Herman
authors.affiliation: h Formerly at Bayer S.A.S., Bayer CropScience , Sophia Antipolis , France.
authors.forename: Corinne
authors.initials: C
authors.lastname: Herouet-Guicheney
authors.affiliation: i DuPont Haskell Global Centers for Health and Environmental Sciences , Newark , DE , USA.
authors.forename: Gregory S
authors.initials: GS
authors.lastname: Ladics
authors.affiliation: j Syngenta Crop Protection, LLC , Research Triangle Park , NC , USA.
authors.forename: Scott
authors.initials: S
authors.lastname: McClain
authors.affiliation: k Allergy Clinic , Copenhagen University Hospital at Gentofte , Copenhagen , Denmark.
authors.forename: Lars K
authors.initials: LK
authors.lastname: Poulsen
authors.affiliation: l Regulatory Science , Formerly at BASF Plant Science , Research Triangle Park , NC , USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Privalle
authors.affiliation: m Regulatory Division, Product Characterization Center , Formerly at Monsanto Co. , St. Louis , MO , USA.
authors.forename: Jason M
authors.initials: JM
authors.lastname: Ward
authors.affiliation: n Protein Allergenicity Technical Committee , ILSI Health and Environmental Sciences Institute (HESI) , Washington , DC , USA.
authors.forename: Nancy
authors.initials: N
authors.lastname: Doerrer
authors.affiliation: f Bayer S.A.S., Bayer CropScience , Sophia Antipolis , France.
authors.forename: Jean-Baptiste
authors.initials: JB
authors.lastname: Rascle
keywords: allergen; multiplex immunodetection; proteomics; rice grain; two-dimensional difference gel electrophoresis (2D-DIGE)