The non-negligible non-specific adsorption of oligonucleotides in target-immobilized Mag-SELEX

文献类型： 外文期刊

第一作者： Liao, Xiaoyan

作者： Liao, Xiaoyan;Huang, Liujuan;Pu, Chunmin;Li, Song;Feng, Bo;Bai, Yalong;Liao, Xiaoyan;Liao, Xiaoyan;Pu, Chunmin;Li, Song

作者机构：

关键词： Mag-SELEX; Non-specific adsorption; Oligonucleotides

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：7.7； 五年影响因子：7.7 ）

ISSN： 0141-8130

年卷期： 2024 年 275 卷

页码：

收录情况： SCI

摘要： Target-immobilized magnetic beads-based Systematic Evolution of Ligands by Exponential Enrichment (targetimmobilized Mag-SELEX) has emerged as a powerful tool for aptamer selection owing to its convenience, efficiency, and versatility. However, in this study we systematically investigated non-specific adsorption in targetimmobilized Mag-SELEX and found that the non-specific adsorption of the oligonucleotides to target-labeled magnetic beads was comparable to that of the screening libraries, indicating a substantial portion of captured sequences likely stem from non-specific adsorption. Longer nucleic acid sequences (80 nt and above, such as polyA80 and yeast tRNA) were found to attenuate this non-specific adsorption, with more complex higher-order structures demonstrating greater efficacy, while dNTP and short sequences such as primer sequences (20 nt), polyT(59), or polyA(59), did not possess this capability. Various evidence suggested that hydrophobic interactions and other weak interactions may be the primary underlying cause of non-specific adsorption. Additionally, surface modification of magnetic beads with polar molecule polyethylene glycol (PEG) also yielded a significant reduction in non-specific adsorption. In conclusion, our research underscores the critical importance of closely monitoring non-specific adsorption in target-immobilized Mag-SELEX.

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