Protein detection using proximity-dependent DNA ligation assays
Protein detection using proximity-dependent DNA ligation assays
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DOI：
10.1038/nbt0502-473
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发表时间：
2002-05-01
影响因子：
46.9
通讯作者：
Landegren, U
中科院分区：
文献类型：
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作者：
Fredriksson, S;Gullberg, M;Landegren, U
体外DNA扩增的出现使得能够快速获取基因组信息。我们在这里提出了一种类似的技术，用于蛋白质检测，其中两个DNA适体的靶蛋白的协调和近端结合促进连接到每个适体亲和探针的寡核苷酸的连接。两个这样的邻近探针的连接产生反映靶蛋白的身份和量的可扩增的DNA序列。这种邻位连接试验检测zeptomole（40 × 10（-21）mol）量的细胞因子血小板衍生生长因子（PDGF），无需洗涤或分离，该机制可以推广到其他形式的蛋白质分析。
The advent of in vitro DNA amplification has enabled rapid acquisition of genomic information. We present here an analogous technique for protein detection, in which the coordinated and proximal binding of a target protein by two DNA aptamers promotes ligation of oligonucleotides linked to each aptamer affinity probe. The ligation of two such proximity probes gives rise to an amplifiable DNA sequence that reflects the identity and amount of the target protein. This proximity ligation assay detects zeptomole (40 x 10(-21) mol) amounts of the cytokine platelet-derived growth factor (PDGF) without washes or separations, and the mechanism can be generalized to other forms of protein analysis.