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The discovery of lactoferrin dual aptamers through surface plasmon resonance imaging combined with a bioinformation analysis.
Analyst ( IF 3.6 ) Pub Date : 2020-09-03 , DOI: 10.1039/d0an01513j Wenchao Jia 1 , Zecheng Wang , Zhongyi Lu , Baiwen Ding , Zhoumin Li , Danke Xu
Analyst ( IF 3.6 ) Pub Date : 2020-09-03 , DOI: 10.1039/d0an01513j Wenchao Jia 1 , Zecheng Wang , Zhongyi Lu , Baiwen Ding , Zhoumin Li , Danke Xu
Affiliation
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An analytical method for screening aptamers for different recognition sites in lactoferrin (Lac) molecules has been developed based on Surface Plasmon Resonance imaging (SPRi), combined with the cluster classification calculation of a quasi-aptamer library strategy and molecular docking simulation analysis. Using the software simulation, a homology analysis was performed on the selected quasi-aptamer sequences, which could be divided into 8 different families. Based on the principle of biomolecular recognition, a label-free, high-throughput dual immune site screening method was established, in which the nucleic acid aptamers of recognizing ability for lactoferrin molecules were fixed onto the surface of the SPRi sensor chip and could bind to the lactoferrin molecules. Then, the aptamer candidates to be paired were introduced, and the recognition event of the second immune site was judged by observing the binding signal of SPRi. The paired SPRi signal was generated only when the site identified by the second nucleic acid molecule was different from the first immune site. Based on this principle, a pair of Lac nucleic acid aptamers (Lac-8 and Lac-25) was finally screened and confirmed using computerized simulation, and has been employed to assay Lac in milk by ELONA (Enzyme-Linked Oligonucleotide Assay).
中文翻译：
通过表面等离振子共振成像结合生物信息分析发现乳铁蛋白双适体。
基于表面等离子体共振成像（SPRi），结合准适体文库策略的聚类分类计算和分子对接模拟分析，开发了一种筛选乳铁蛋白（Lac）分子中不同识别位点的适体的分析方法。使用软件模拟，对所选的准适体序列进行同源性分析，该序列可分为8个不同的家族。基于生物分子识别原理，建立了一种无标记，高通量双重免疫部位筛选方法，该方法将对乳铁蛋白分子具有识别能力的核酸适体固定在SPRi传感器芯片的表面上并可以与乳铁蛋白分子。然后，介绍了要配对的适体候选者，通过观察SPRi的结合信号判断第二免疫位点的识别事件。仅当第二核酸分子识别的位点与第一免疫位点不同时才生成配对的SPRi信号。基于此原理，最终使用计算机模拟筛选并确认了一对Lac核酸适体（Lac-8和Lac-25），并已通过ELONA（酶联寡核苷酸测定）用于牛奶中的Lac测定。
更新日期：2020-09-28
中文翻译：
通过表面等离振子共振成像结合生物信息分析发现乳铁蛋白双适体。
基于表面等离子体共振成像（SPRi），结合准适体文库策略的聚类分类计算和分子对接模拟分析，开发了一种筛选乳铁蛋白（Lac）分子中不同识别位点的适体的分析方法。使用软件模拟，对所选的准适体序列进行同源性分析，该序列可分为8个不同的家族。基于生物分子识别原理，建立了一种无标记，高通量双重免疫部位筛选方法，该方法将对乳铁蛋白分子具有识别能力的核酸适体固定在SPRi传感器芯片的表面上并可以与乳铁蛋白分子。然后，介绍了要配对的适体候选者，通过观察SPRi的结合信号判断第二免疫位点的识别事件。仅当第二核酸分子识别的位点与第一免疫位点不同时才生成配对的SPRi信号。基于此原理，最终使用计算机模拟筛选并确认了一对Lac核酸适体（Lac-8和Lac-25），并已通过ELONA（酶联寡核苷酸测定）用于牛奶中的Lac测定。
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