Swiss Institute of Allergy and Asthma Research (SIAF), University of Zurich, Davos, Switzerland.
Research Center for Translational Medicine (KUTTAM), Koç University School of Medicine, Istanbul, Turkey.
Pediatr Allergy Immunol. 2024 Jun;35(6):e14174. doi: 10.1111/pai.14174.
Milk oral immunotherapy is the riskiest and most unpredictable form of oral immunotherapy. We aimed to produce a low allergenic product than conventional once baked-cake/muffin, to develop indirect in-house ELISA to check the tolerance status with milk products and evaluate IgE reactivity of patients' sera via western blotting (WB) and indirect in-house ELISA.
A low allergenic product named biscotti-twice baked-cake was developed, and the total protein concentration was determined. The protein content was studied by SDS-PAGE and proteomics. Milk-specific IgE (sIgE) binding assays were performed by WB and indirect in-house ELISA by using patients' sera.
Casein band intensity was observed to be lower in the biscotti-twice baked-cake than in the once baked-cake (p = .014). Proteomics analysis and αS1-casein measurement showed that the lowest intensity of casein was found in biscotti. The low binding capacity of milk sIgE to biscotti compared with once baked-cake was shown by WB (p = .0012) and by indirect in-house ELISA (p = .0001). In the ROC analysis, the area under the curve (AUC) of the in-house ELISA IgE was comparable with Uni-CAP milk and casein sIgE. The AUC of the in-house ELISA IgE for cake (0.96) and biscotti (1) was slightly better than Uni-CAP milk sIgE (0.94; 0.97) and casein sIgE (0.96; 0.97), respectively.
The low allergenicity of the newly developed low allergenic product "biscotti-twice baked-cake" has been demonstrated by in vitro experiments. Biscotti could be a safe treatment option than once baked-cake/muffin in patients who are reactive to once baked-milk products.
牛奶口服免疫疗法是风险最高、最不可预测的口服免疫疗法形式。我们旨在生产出一种比传统的一次烘焙蛋糕/松饼过敏原更低的产品，开发间接内部 ELISA 来检查牛奶产品的耐受状态，并通过 Western blot (WB) 和间接内部 ELISA 评估患者血清中的 IgE 反应性。
开发了一种名为 biscotti-二次烘焙蛋糕的低过敏原产品，并确定了总蛋白浓度。通过 SDS-PAGE 和蛋白质组学研究蛋白质含量。通过 WB 和间接内部 ELISA 用患者血清进行牛奶特异性 IgE (sIgE) 结合测定。
与一次烘焙蛋糕相比， biscotti-二次烘焙蛋糕中的酪蛋白条带强度较低（p=0.014）。蛋白质组学分析和 αS1-酪蛋白测量显示， biscotti 中的酪蛋白强度最低。WB（p=0.0012）和间接内部 ELISA（p=0.0001）显示，与一次烘焙蛋糕相比，牛奶 sIgE 对 biscotti 的结合能力较低。在 ROC 分析中，内部 ELISA IgE 的曲线下面积（AUC）与 Uni-CAP 牛奶和酪蛋白 sIgE 相当。内部 ELISA IgE 对蛋糕（0.96）和 biscotti（1）的 AUC 略优于 Uni-CAP 牛奶 sIgE（0.94；0.97）和酪蛋白 sIgE（0.96；0.97）。
通过体外实验证明了新开发的低过敏原产品“biscotti-二次烘焙蛋糕”的低过敏原性。与一次烘焙的牛奶产品反应的患者， biscotti 可能是一种比一次烘焙蛋糕/松饼更安全的治疗选择。
