BACKGROUND The commercial wheat extract for skin prick test (SPT) provides less sensitivity to predict wheat allergy, compared to in-house gliadin extracts. SPT is a preferred method to study extract stability as it is the aim of developing extract. The role of cell degranulation assay, a functional assay with the same mechanism as SPT, is not widely used to determine extract stability. OBJECTIVE To study the stability of in-house gliadin extracts stored at different periods, by using protein analysis, SPT and degranulation assay of humanized rat basophilic-leukemia (RBL-SX38) cells. METHODS Patients with a history of wheat allergy and positive SPT to wheat, were recruited. The gliadin extracts stored for 1, 6, 9, and 12 months at 2-8°C were used in SDS-PAGE, SPT and cell degranulation assay. The cell degranulation was determined by β-hexosaminidase release. AR patients. RESULTS Forty children were recruited. The gliadin extract stored for 9 and 12 months provided lighter protein bands than 1 and 6 months. However, the wheal diameters from SPT using extracts stored at different periods, were not significantly different (p = 0.09). There were also no significant differences of the β-hexosaminidase released using 0.1 and 1 μg/mL of gliadin extracts stored at different periods (p > 0.05). The 10 μg/mL of gliadin extracts stored at longer periods, significantly stimulated higher β-hexosaminidase release (p = 0.01). The extracts were sterile at all storage times. CONCLUSIONS To determine the stability of in-house gliadin extracts, SPT or cell degranulation assay provided additional information to SDS-PAGE. The extracts were stable for up to 12 months.

中文翻译：

---

通过蛋白质分析、皮肤测试和细胞脱粒测定来鉴定新型小麦麦醇溶蛋白提取物的稳定性。

背景与内部麦醇溶蛋白提取物相比，用于皮肤点刺试验（SPT）的商业小麦提取物在预测小麦过敏方面的敏感性较低。SPT 是研究提取物稳定性的首选方法，因为它是开发提取物的目的。细胞脱粒测定是一种与 SPT 机制相同的功能测定，其作用并未广泛用于确定提取物的稳定性。目的通过对人源化大鼠嗜碱性白血病（RBL-SX38）细胞进行蛋白质分析、SPT和脱粒实验，研究不同时期保存的内部麦醇溶蛋白提取物的稳定性。方法 招募有小麦过敏史且小麦 SPT 阳性的患者。将在2-8°C下储存1、6、9和12个月的麦醇溶蛋白提取物用于SDS-PAGE、SPT和细胞脱颗粒测定。通过β-己糖胺酶的释放来测定细胞脱颗粒。AR患者。结果 四十名儿童被招募。储存 9 个月和 12 个月的麦醇溶蛋白提取物提供的蛋白质条带比储存 1 个月和 6 个月的蛋白条带更浅。然而，使用不同时期储存的提取物进行的 SPT 的风团直径没有显着差异 (p = 0.09)。使用0.1和1μg/mL的麦醇溶蛋白提取物在不同时期储存时释放的β-己糖胺酶也没有显着差异（p>0.05）。10 μg/mL 麦醇溶蛋白提取物储存时间较长，可显着刺激更高的 β-己糖胺酶释放 (p = 0.01)。提取物在所有储存时间都是无菌的。结论 为了确定内部麦醇溶蛋白提取物的稳定性，SPT 或细胞脱颗粒测定为 SDS-PAGE 提供了额外的信息。提取物可稳定保存长达 12 个月。