Epitope map of two polyclonal antibodies that recognize amyloid lesions in patients with Alzheimer's disease.
Epitope map of two polyclonal antibodies that recognize amyloid lesions in patients with Alzheimer's disease.
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识别阿尔茨海默病患者淀粉样蛋白病变的两种多克隆抗体的表位图。
DOI：
10.1042/bj2820517
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Frangione,B
中科院分区：
文献类型：
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作者：
Ghiso,J;Wisniewski,T;Vidal,R;Rostagno,A;Frangione,B
两种合成肽的序列与阿尔茨海默病淀粉样前体蛋白 (APP) 胞外结构域片段的序列相同，用于产生抗体。 SP28 包含 APP695 同工型的位置 597-624，而 SP41 向 N 末端延伸（氨基酸 584-624）并包含整个 SP28 肽。使用 e.l.i.s.a.和抑制实验中，我们鉴定出两个含有β转角的片段602-607和617-624分别作为抗SP41和抗SP28识别的表位。这两种抗体都对阿尔茨海默病患者和患有相关疾病的患者大脑中的淀粉样蛋白病变进行了免疫标记，而在对照大脑中则没有反应。然而，当用免疫印迹检测时，抗SP28未能从杆状病毒感染的Sf9细胞中检测到全长APP，并且与其他抗APP抗血清相比，抗SP41反应较弱。数据表明，这些抗体针对天然分子中不存在但在替代 APP 处理后存在的构象表位。
Two synthetic peptides with sequences identical with those of fragments of the extracellular domain of the Alzheimer's-disease amyloid precursor protein (APP) were used to raise antibodies. SP28 comprises positions 597-624 of the APP695 isoform, whereas SP41 extends towards the N-terminus (amino acids 584-624) and contains the entire SP28 peptide. Using e.l.i.s.a. and inhibition experiments we identified the two beta-turn-containing segments 602-607 and 617-624 as the epitopes recognized by anti-SP41 and anti-SP28 respectively. Both antibodies immunolabelled amyloid lesions in brains from Alzheimer's-disease patients and patients with related disorders, whereas they were unreactive in control brains. However, when probed on immunoblots, anti-SP28 failed to detect full-length APP from baculovirus-infected Sf9 cells, and anti-SP41 reacted weakly compared with other anti-APP antisera. The data suggest that these antibodies are directed to conformational epitopes not existent in the native molecules but present after alternative APP processing.