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目的筛选粉尘螨(Der f)主要和中等效价变应原(Der f1、Der f2、Der f4、Der f5和Der f7)可能具有的线性表位。方法根据Der f1、Der f2、Der f4、Der f5和Der f7活性部位氨基酸序列,各合成含8个氨基酸的短肽,相互之间重叠7个氨基酸,将这些短肽点到芯片上构建微阵列,将此微阵列芯片与人血清Ig E进行免疫标记反应,扫描芯片并分析数据。结果合成了5组变应原共1128个短肽,并构建微阵列芯片。将6份对粉尘螨过敏的儿童血清混合并加样到微阵列芯片上,进行扫描和数据分析,最终得到Der f1的4个表位(第46~53位氨基酸、第71~78位氨基酸、第99~110位氨基酸和第179~186位氨基酸),Der f2的3个表位(第15~22位氨基酸、第80~89位氨基酸和第106~113位氨基酸),Der f4的6个表位(第69~82位氨基酸、第107~116位氨基酸、第225~232位氨基酸、第261~268位氨基酸、第355~365位氨基酸和第483~496位氨基酸),Der f5的1个表位(第102~109位氨基酸)和Der f7的3个表位(第32~39位氨基酸,第52~64位氨基酸和第100~107位氨基酸)。结论确定了粉尘螨变应原Der f1、Der f2、Der f4、Der f5和Der f7线性表位。
Objective To screen for possible linear epitopes of Der f1, Der f2, Der f4, Der f5 and Der f7. Methods Based on the amino acid sequences of Der f1, Der f2, Der f4, Der f5 and Der f7, 8-amino-acid short peptides were synthesized and overlapped with each other by 7 amino acids. These short peptides were spotted on chip to construct microarray , This microarray chip and human serum Ig E immunolabeling reaction, scanning the chip and analyze the data. Results A total of 1128 short peptides of 5 allergens were synthesized and microarray chips were constructed. Six serums of children allergic to dust mites were mixed and loaded onto microarray chips for scanning and data analysis to finally obtain four epitopes of Der f1 (amino acids 46-53, amino acids 71-78, Amino acids 99 to 110, and amino acids 179 to 186), three epitopes of Der f2 (amino acids 15 to 22, amino acids 80 to 89, and amino acids 106 to 113), six of Der f4 Epitopes (amino acids 69-82, amino acids 107-116, amino acids 225-232, amino acids 261-268, amino acids 355-365, and amino acids 483-496), 1 (Epitopes 102-109) and Der f7 (amino acids 32-39, amino acids 52-64 and amino acids 100-107) of Der f7. Conclusion The linear epitopes of Der f1, Der f2, Der f4, Der f5 and Der f7 of the dust mite allergens were identified.