Beaugrand Johnny, Gebruers Kurt, Ververken Cedric, Fierens Ellen, Dornez Emmie, Goddeeris Bruno M, Delcour Jan A, Courtin Christophe M
Laboratory of Food Chemistry and Biochemistry, Katholieke Universiteit Leuven, Kasteelpark Arenberg 30, B-3001 Leuven, Belgium.
J Agric Food Chem. 2007 Sep 19;55(19):7682-8. doi: 10.1021/jf071087b. Epub 2007 Aug 24.
To quantify Triticum aestivum xylanase inhibitor (TAXI) and xylanase inhibiting protein (XIP) type proteins in cereals in general and wheat ( T. aestivum) in particular, a robust enzyme-linked immunosorbent assay (ELISA) using an uncommon enzyme-antibody sandwich format was developed. Bacillus subtilis glycoside hydrolase family (GH) 11 and Aspergillus oryzae GH 10 xylanases were selected for coating ELISA plate wells to capture TAXI and XIP, respectively, prior to probing with antibodies. The detection threshold of the developed ELISA was much lower than that of the currently used xylanase inhibitor assay and the recently described Western blot approach. Because of its broad dynamic range (TAXI, 30-600 ng/mL, and XIP, 3-60 ng/mL), one proper standard extract dilution can be used for analyzing different wheat varieties, whereas for the currently used colorimetric assay, often different dilutions need to be analyzed. The TAXI ELISA for wheat was successfully adapted for barley ( Hordeum vulgare) and could also be used for other cereals.
为了定量分析谷物尤其是小麦（普通小麦）中的小麦木聚糖酶抑制剂（TAXI）和木聚糖酶抑制蛋白（XIP）类型的蛋白质，开发了一种使用罕见的酶 - 抗体夹心形式的稳健酶联免疫吸附测定（ELISA）方法。选择枯草芽孢杆菌糖苷水解酶家族（GH）11木聚糖酶和米曲霉GH 10木聚糖酶分别包被ELISA板孔，用于捕获TAXI和XIP，然后用抗体进行检测。所开发的ELISA的检测阈值远低于目前使用的木聚糖酶抑制剂测定法和最近描述的蛋白质印迹法。由于其宽动态范围（TAXI为30 - 600 ng/mL，XIP为3 - 60 ng/mL），一种合适的标准提取物稀释液可用于分析不同的小麦品种，而对于目前使用的比色测定法，通常需要分析不同的稀释度。用于小麦的TAXI ELISA已成功适用于大麦（大麦），也可用于其他谷物。
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