Legen Tjasa, Mayer Günter
Life and Medical Sciences Institute (LIMES), University of Bonn, 53121, Germany.
Center of Aptamer Research and Development (CARD), University of Bonn, 53121, Germany.
Chembiochem. 2025 Aug 22;26(15):e202500264. doi: 10.1002/cbic.202500264. Epub 2025 Jul 9.
Due to their small size, stability, and cost-effectiveness compared to antibodies, aptamers developed by systematic evolution of ligands by exponential enrichment (SELEX) are promising candidates for the detection of small molecules. In SELEX, a small target molecule is usually covalently immobilized on a surface to separate bound from unbound nucleic acid sequences. However, this immobilization alters the molecule, that is, additional chemical entities are added and the electron distribution is altered, compromising the enrichment properties. To overcome this problem, a capture SELEX method has been successfully developed in which the RNA or DNA libraries are bound to a surface via a complementary oligodeoxynucleotide, and the soluble ligand is used to capture nucleic acids that bind to it from that surface. Herein, the development of an automated version of the capture SELEX method for the identification of RNA aptamers that bind small molecules is described. This method is fully automated and performs up to 12 iterative selection cycles without manual interference in 72 h. The approach is therefore suitable as rapid route to aptamers and enables resource-efficient test selections to assess "aptamerogenicity" of a target.
与抗体相比,通过指数富集的配体系统进化技术(SELEX)开发的适体,因其尺寸小、稳定性高且具有成本效益,是检测小分子的有前景的候选物。在SELEX中,通常将小分子靶标共价固定在表面上,以分离结合的和未结合的核酸序列。然而,这种固定会改变分子,即添加了额外的化学实体并改变了电子分布,从而损害了富集特性。为克服这一问题,已成功开发了一种捕获SELEX方法,其中RNA或DNA文库通过互补的寡脱氧核苷酸与表面结合,可溶性配体用于从该表面捕获与其结合的核酸。本文描述了一种用于鉴定与小分子结合的RNA适体的自动化捕获SELEX方法的开发。该方法完全自动化,可在72小时内无人工干预地执行多达12个迭代选择循环。因此,该方法适合作为获得适体的快速途径,并能够进行资源高效的测试选择,以评估靶标的“适体生成能力”。