Screening specific aptamer pairs towards bovine lactoferrin for its sandwich application

文献类型： 外文期刊

第一作者： Feng, Ziru

作者： Feng, Ziru;Yan, Mengmeng;Zhang, Wenjun;Zhu, Chao;Feng, Ziru;Li, Tengfei

作者机构：

关键词： Lactoferrin; Aptamer; SELEX; Aptamer pairs; Sandwich applications

期刊名称：FOOD BIOSCIENCE （ 影响因子：5.9； 五年影响因子：6.1 ）

ISSN： 2212-4292

年卷期： 2025 年 64 卷

页码：

收录情况： SCI

摘要： Lactoferrin (Lf) is closely associated with the growth of infants and children, and its recognition is imperative due to its rising application as a prevalent nutritional fortification in infant formula. In this study, an ssDNA aptamer was screened out that enabled sandwich recognition of Lf by forming an aptamer pair with another available aptamer. During the process, the reported aptamer (SeqT1.1) was selected as the substrateto be linked onto pre- blocked microplates for capturing Lf, followed by the addition of the ssDNA library, resulting in the formation of the sandwich complex of the "SeqT1.1/Lf/ssDNA library" that was subsequently collected for screening. After seven rounds, we discovered a new ssDNA aptamer (Seq6T) through a comprehensive analysis involving sequence frequency, secondary structures, homology, and sequence truncation, which helped address the binding interference between aptamer pairs with each other's binding. The Seq6T demonstrated good specificity and lownanomolar affinity by ELONA (KD, 12.59 nM) and CE-LIF (KD, 22.44 nM) assays, which illustrated the potential of applying that in both homogeneous and non-homogeneous environments. The sandwich recognition of "SeqT1.1/Lf/Seq6T" was further verified by molecular docking and ELONA, with a good detection limit of 18.35 nM. The aptamer pairs provided a considerable basis for accelerating the sandwich recognition applications towards Lf.

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