Antibody binding of deletion mutants of Asp f 2, the major Aspergillus fumigatus allergen.
Antibody binding of deletion mutants of Asp f 2, the major Aspergillus fumigatus allergen.
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主要烟曲霉过敏原 Asp f 2 缺失突变体的抗体结合。
DOI：
10.1006/bbrc.2000.2546
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Kurup,VP
中科院分区：
文献类型：
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作者：
Tang,B;Banerjee,B;Greenberger,PA;Fink,JN;Kelly,KJ;Kurup,VP
Aspf 2是烟曲霉（Aspergillusfumigatus，Af）的主要变应原，由268个氨基酸组成，具有9个线性IgE结合区.不知道这些线性表位中的任何一个是否也是适形表位。因此，我们构建了缺失一个或多个表位的Aspf 2缺失突变体，并将这些蛋白与ABPA患者血清的IgE结合与在E. coli和Pichia。与E.大肠杆菌表达Aspf 2。当C-末端或N-末端缺失时，仅观察到弱IgE结合，而两端的耗尽否定了所有反应性。单克隆抗体IL-B8和ABPA血清中的IgE和IgG与Aspf 2 E-4片段显著结合，表明主要的B细胞表位位于Aspf 2的N-末端。
Asp f 2, a 268 amino acid major allergen from Aspergillus fumigatus (Af) demonstrated nine linear IgE binding regions. It is not known whether any of these linear epitopes are also conformatory epitopes. Hence, we constructed deletion mutants of Asp f 2 devoid of one or more epitopes, and the IgE binding of these proteins with sera from patients with ABPA was compared with the full-length Asp f 2 expressed in E. coli and Pichia. The Pichia expressed protein reacted weakly with IgE, but strongly with IgG of ABPA sera compared to E. coli expressed Asp f 2. Weak IgE binding only was seen when the C-terminal or N-terminal was deleted, while depletion of both ends negated all reactivity. The monoclonal antibody IL-B8 and IgE and IgG of ABPA sera bound significantly to the Asp f 2 E-4 fragment indicating that the major B-cell epitope is located at the N-terminal end of Asp f 2.