Department of Applied Chemistry, Graduate School of Engineering, Osaka Metropolitan University, Osaka, Japan.
CREST, Japan Science and Technology Agency, Tokyo, Japan.
Anal Sci. 2024 Aug;40(8):1499-1508. doi: 10.1007/s44211-024-00588-6. Epub 2024 Jun 11.
In this study, a structure-induced aptamer targeting small molecules was selected using capillary sieving electrophoresis (CSE). CSE was conducted using a capillary filled with a background solution containing hydroxypropyl cellulose as a sieving matrix to separate the aptamer candidates by changing their structures via complexation. Before aptamer selection, the original random-sequence DNA library was used to create structure-not-preorganized DNA sub-library containing straight-chain-like structures using CSE. Next, a structure-induced aptamer targeting L-tyrosinamide was selected from the prepared sub-library. Six aptamer candidates were selected, one of which showed a binding ability comparable to that of the reported L-tyrosinamide aptamer and selectivity toward the analogs. These results indicated that the proposed method can be applied to select structure-induced aptamers that target small molecules.
在这项研究中，使用毛细管筛分电泳（CSE）选择了一种针对小分子的结构诱导适体。CSE 通过使用充满包含羟丙基纤维素的背景溶液的毛细管进行，通过络合改变适体候选物的结构来对其进行分离。在适体选择之前，原始随机序列 DNA 文库被用于使用 CSE 创建包含直链样结构的未预组织 DNA 亚文库。接下来，从制备的亚文库中选择了一种针对 L-酪氨酸酰胺的结构诱导适体。从六个适体候选物中选择了一个，它表现出与报道的 L-酪氨酸酰胺适体相当的结合能力，并对类似物具有选择性。这些结果表明，所提出的方法可用于选择针对小分子的结构诱导适体。
