Division of Molecular Therapeutics and Formulation, School of Pharmacy, University of Nottingham, Nottingham, UK.
School of Biosciences, University of Nottingham, Nottingham, UK.
Methods Mol Biol. 2020;2163:155-162. doi: 10.1007/978-1-0716-0696-4_12.
The presence of allergen-specific IgE (sIgE) in human sera can be determined by measuring the binding of sIgE to solid phase-bound preparations containing the allergens to be tested. These can be complex extracts, purified or recombinant allergens, or peptides. Older methods, such as the IgE CAP test, only allow sIgE measurements to multiple allergens in individual measurements. Newer technologies such as the ImmunoCAP ISAC test allows semiquantitative testing of sIgE to over a hundred allergens on a protein array. Allergen arrays have higher numerical power, allowing testing to many allergens at the same time, using only a small amount of serum. We have previously demonstrated how allergen arrays can be used in combination with purified peripheral blood basophils, introducing a clinically relevant readout. Here, we describe a protocol and materials that allow the testing of sIgE with multiple allergens in array format, using a humanized fluorescent IgE reporter system (RBL NFAT-DsRed).
人血清中过敏原特异性 IgE（sIgE）的存在可以通过测量 sIgE 与包含待测试过敏原的固相制剂的结合来确定。这些制剂可以是复杂的提取物、纯化或重组过敏原，或肽。较旧的方法，如 IgE CAP 测试，仅允许在单个测量中测量多种过敏原的 sIgE。更新的技术，如 ImmunoCAP ISAC 测试，允许在蛋白质阵列上对半定量测试超过 100 种过敏原的 sIgE。过敏原阵列具有更高的数值能力，允许同时使用少量血清对许多过敏原进行测试。我们之前已经证明了如何将过敏原阵列与纯化的外周血嗜碱性粒细胞结合使用，引入了一种临床相关的读出方法。在这里，我们描述了一种使用人源化荧光 IgE 报告系统（RBL NFAT-DsRed）以阵列格式测试多种过敏原的 sIgE 的方案和材料。
