ISOLATION AND QUANTIFICATION OF SOLUBLE ALZHEIMERS BETA-PEPTIDE FROM BIOLOGICAL-FLUIDS
ISOLATION AND QUANTIFICATION OF SOLUBLE ALZHEIMERS BETA-PEPTIDE FROM BIOLOGICAL-FLUIDS
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DOI：
10.1038/359325a0
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发表时间：
1992-09-24
期刊：
影响因子：
64.8
通讯作者：
SCHENK, D
中科院分区：
文献类型：
--
作者：
SEUBERT, P;VIGOPELFREY, C;SCHENK, D
β - 淀粉样肽（A - β）在脑内的沉积是阿尔茨海默病的一个不变特征。自从A - β最初被分离和鉴定（参考文献1）以及其前体蛋白随后被克隆（参考文献2 - 5）以来，尚未有关于离散A - β实际产生的直接证据被报道（参考文献6 - 11）。在此我们利用针对A - β内一个跨越正常组成性切割位点的表位的特异性抗体，研究A - β是否存在于人体生物体液中（参考文献12，13）。这些抗体被用于构建一种夹心式酶联免疫吸附测定法，该方法可检测脑脊液、血浆以及体外培养的人混合脑细胞的条件培养基中的A - β（另见参考文献14）。通过亲和层析，我们从人脑脊液和人混合脑细胞培养的条件培养基中纯化并测定了A - β以及一种新的A - β片段的序列。这些发现表明A - β在体内和体外均可产生和释放。这些观察结果为开发阿尔茨海默病的诊断测试以及旨在减少A - β在脑内沉积的治疗策略提供了新的契机。
CEREBRAL deposition of the beta-amyloid peptide (A-beta) is an invariant feature of Alzheimer's disease. Since the original isolation and characterization of A-beta (ref. 1) and the subsequent cloning of its precursor protein2-5, no direct evidence for the actual production of discrete A-beta has been reported6-11. Here we investigate whether A-beta is present in human biological fluids using antibodies specific for an epitope within A-beta that spans the site of normal constitutive cleavage12,13. These antibodies were used to construct a sandwich-type enzyme-linked immunosorbent assay that detects A-beta in cerebrospinal fluid, plasma and conditioned medium of human mixed-brain cells grown in vitro (see also ref. 14). By affinity chromatography, we have purified and sequenced A-beta and a novel A-beta fragment from human cerebrospinal fluid and conditioned medium of human mixed-brain cell cultures. These findings demonstrate that A-beta is produced and released both in vivo and in vitro. These observations offer new opportunities for developing diagnostic tests for Alzheimer's disease and therapeutic strategies aimed at reducing the cerebral deposition of A-beta.