A modified extraction protocol enables detection and quantification of celiac disease-related gluten proteins from wheat.
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.
van den Broeck
Hetty C
HC
Plant Research International, Wageningen UR, PO Box 16, 6700 AA Wageningen, The Netherlands. hetty.busink@wur.nl
America
Antoine H P
AH
Smulders
Marinus J M
MJ
Bosch
Dirk
D
Hamer
Rob J
RJ
Gilissen
Ludovicus J W J
LJ
van der Meer
Ingrid M
IM
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
D052179:Antigens, Plant; D010936:Plant Extracts; D005983:Glutens