Haas-Lauterbach Sigrid, Immer Ulrike, Richter Mareike, Koehler Peter
R-Biopharm, An der neuen Bergstrasse 17, D-64297 Darmstadt, Germany.
J AOAC Int. 2012 Mar-Apr;95(2):377-81. doi: 10.5740/jaoacint.sge_haas-lauterbach.
The second generation of a competitive ELISA for prolamin quantification based on the R5 antibody was studied for method performance and suitability to detect partially hydrolyzed prolamins in food. To be able to convert signal intensities to gluten concentrations, as required by the Codex Alimentarius Standard, a new calibrator consisting of a peptic-tryptic digest of wheat, rye, and barley prolamins was used for the first time. LOD and LOQ of the assay were 1.36 and 5.0 mg prolamin/kg food, respectively. Analysis of beer samples and a hydrolyzed wheat product showed that the assay provided significantly higher prolamin concentrations, compared to the sandwich ELISA based on the same antibody, which is only suitable for the detection of intact prolamins. Spiking experiments with defined concentrations of partially hydrolyzed prolamins gave recoveries ranging from 92 to 136%.
研究了基于R5抗体的用于醇溶蛋白定量的第二代竞争性酶联免疫吸附测定法（ELISA）的方法性能，以及该方法检测食品中部分水解醇溶蛋白的适用性。为了能够按照食品法典标准的要求将信号强度转换为面筋浓度，首次使用了一种由小麦、黑麦和大麦醇溶蛋白的胃蛋白酶-胰蛋白酶消化物组成的新型校准物。该测定法的检测限（LOD）和定量限（LOQ）分别为1.36和5.0毫克醇溶蛋白/千克食品。对啤酒样品和一种水解小麦产品的分析表明，与仅适用于检测完整醇溶蛋白的基于相同抗体的夹心ELISA相比，该测定法提供的醇溶蛋白浓度显著更高。用确定浓度的部分水解醇溶蛋白进行的加标实验回收率在92%至136%之间。
