Selection of Aptamers Against Whole Living Cells: From Cell-SELEX to Identification of Biomarkers
Selection of Aptamers Against Whole Living Cells: From Cell-SELEX to Identification of Biomarkers
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DOI：
10.1007/978-1-4939-6857-2_16
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发表时间：
2017-01-01
期刊：
影响因子：
--
通讯作者：
Duconge, Frederic
中科院分区：
文献类型：
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作者：
Nam Nguyen Quang;Miodek, Anna;Duconge, Frederic
适体选择方案（称为 cell-SELEX）已开发用于使用整个活细胞作为靶标来靶向跨膜蛋白。该技术具有多个优点。 (1)不需要使用纯化的蛋白质。 (2)针对处于其天然构象的膜蛋白选择适体。 (3) 可以进行 Cell-SELEX 来识别针对不同细胞系之间差异表达的生物标志物的适体，而无需事先了解目标。 (4) cell-SELEX鉴定的适体可进一步用于纯化其靶标并鉴定新的生物标志物。在这里，我们提供了 cell-SELEX 方案，包括寡核苷酸文库的制备、下一代测序和放射性结合测定。此外，我们还提供了纯化和鉴定这些适体目标的方案。这些方案可用于发现先导治疗化合物和诊断细胞表面生物标志物。
Aptamer selection protocols, named cell-SELEX, have been developed to target trans-membrane proteins using whole living cells as target. This technique presents several advantages. (1) It does not necessitate the use of purified proteins. (2) Aptamers are selected against membrane proteins in their native conformation. (3) Cell-SELEX can be performed to identify aptamers against biomarkers differentially expressed between different cell lines without prior knowledge of the targets. (4) Aptamers identified by cell-SELEX can be further used to purify their targets and to identify new biomarkers. Here, we provide a protocol of cell-SELEX including the preparation of an oligonucleotide library, next-generation sequencing and radioactive binding assays. Furthermore, we also provide a protocol to purify and identify the target of these aptamers. These protocols could be useful for the discovery of lead therapeutic compounds and diagnostic cell-surface biomarkers.