应用短肽阵列筛选哮喘患儿血清粉尘螨变应原特异性IgE抗体的结合表位

发布时间：2018-06-01 16:58

  本文选题：尘螨变应原 + 肽微阵列　； 参考：《细胞与分子免疫学杂志》2017年08期

【摘要】：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。
[Abstract]:Objective to screen the possible linear epitopes of Der f5 and Der f7 in Der f1, Der f2, Der f4, Der f5 and Der f7, the main and middle titer allergens of Dermatophagoides farinae. Methods according to the amino acid sequences of the active sites of Der f1, Der f2, Der f4, Der f5 and Der f7, each short peptide containing 8 amino acids was synthesized, and 7 amino acids were overlapped with each other. These short peptides were placed on the chip to construct a microarray. The microarray was immunoreactive with human serum IgE, the microarray was scanned and the data were analyzed. Results 1128 short peptides were synthesized from 5 groups of allergens and microarray chips were constructed. Six sera of children allergic to Dermatophagoides farinae were mixed and sampled on microarray chip. The four epitopes of Der F1 were obtained by scanning and data analysis. The three epitopes of the 99th, 110th, 179-, 186th, 225232, 261-, 261-, 89th, 106-, 113amino acids and Der f4 epitopes of Der f4, 69,82,107-, 225232, 261-, 261-, respectively, were found at the 99th, 110th, 186th, 225232, 261-, 261-, 106-, 106-, 113th, 107-, 116th, 225232, 261- and 261268 amino acids, respectively. One epitope of Der f5 (102109amino acid) and three epitopes of Der f7 (32h3t 39-amino acid, 52r-64-amino acid and 100,107amino acid) were found at 355C 365 amino acid and 483H 496 amino acid Der f5. Conclusion the linear epitopes of Der f 2, Der f 4, Der f 5 and Der f 7 of the allergen Der f 1, Der f 2, Der f 4, were identified.
【作者单位】： 江苏医药职业学院基础医学部;盐城市第三人民医院检验科;
【基金】：国家自然科学基金(31272369)
【分类号】：R725.6
，

本文编号：1964971