title: A modified extraction protocol enables detection and quantification of celiac disease-related gluten proteins from wheat.
abstract: The detection, analysis, and quantification of individual celiac disease (CD) immune responsive gluten proteins in wheat and related cereals (barley, rye) require an adequate and reliable extraction protocol. Because different types of gluten proteins behave differently in terms of solubility, currently different extraction protocols exist. The performance of various documented gluten extraction protocols is evaluated for specificity and completeness by gel electrophoresis (SDS-PAGE), immunoblotting and RIDASCREEN Gliadin competitive ELISA. Based on these results, an optimized, two-step extraction protocol has been developed.
authors.affiliation: Plant Research International, Wageningen UR, PO Box 16, 6700 AA Wageningen, The Netherlands. hetty.busink@wur.nl
authors.forename: Hetty C
authors.initials: HC
authors.lastname: van den Broeck
authors.forename: Antoine H P
authors.initials: AH
authors.lastname: America
authors.forename: Marinus J M
authors.initials: MJ
authors.lastname: Smulders
authors.forename: Dirk
authors.initials: D
authors.lastname: Bosch
authors.forename: Rob J
authors.initials: RJ
authors.lastname: Hamer
authors.forename: Ludovicus J W J
authors.initials: LJ
authors.lastname: Gilissen
authors.forename: Ingrid M
authors.initials: IM
authors.lastname: van der Meer
mesh_terms: D052179:Antigens, Plant / Q000737:chemistry* / Q000302:isolation & purification; D002446:Celiac Disease / Q000276:immunology*; D005591:Chemical Fractionation / Q000379:methods*; D005433:Flour / Q000032:analysis; D005983:Glutens / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification; D014908:Triticum / Q000737:chemistry* / Q000276:immunology
chemical_list: D052179:Antigens, Plant; D010936:Plant Extracts; D005983:Glutens