van den Broeck Hetty C, America Antoine H P, Smulders Marinus J M, Bosch Dirk, Hamer Rob J, Gilissen Ludovicus J W J, van der Meer Ingrid M
Plant Research International, Wageningen UR, PO Box 16, 6700 AA Wageningen, The Netherlands.
J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Apr 1;877(10):975-82. doi: 10.1016/j.jchromb.2009.02.035. Epub 2009 Feb 25.
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.
检测、分析和定量小麦及相关谷物(大麦、黑麦)中个体乳糜泻(CD)免疫反应性麸质蛋白需要一个充分且可靠的提取方案。由于不同类型的麸质蛋白在溶解性方面表现不同,目前存在不同的提取方案。通过凝胶电泳(SDS-PAGE)、免疫印迹和RIDASCREEN麦醇溶蛋白竞争性酶联免疫吸附测定,评估各种已记录的麸质提取方案在特异性和完整性方面的性能。基于这些结果,已开发出一种优化的两步提取方案。