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首页> 外文期刊>RSC Advances >Evaluation of aptamer specificity with or without primers using clinical samples for C-reactive protein by magnetic-assisted rapid aptamer selection
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Evaluation of aptamer specificity with or without primers using clinical samples for C-reactive protein by magnetic-assisted rapid aptamer selection

机译：通过磁辅助快速适体选择，使用使用临床样品的临床样品的临床样品进行适体特异性的评估

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摘要

Aptamers with primer binding sites are necessary for the SELEX (Systematic Evolution of Ligands by EXponential enrichment) process. Such primer sequences cause non-specific binding by their nature and raise the incidence of false positives. Thus, the use of aptamers without primer or with minimum primer, to shorten the length of selected aptamers, has become an interesting research topic. Recently, a primer-free aptamer selection protocol based on the magnetic-assisted rapid aptamer selection (MARAS) technology has been reported to generate primer-free aptamers with high affinity and specificity. Adversely, the yield of a suitable aptamer was low and the advantage of MARAS was deteriorated. In this study, multiple negative selection runs were used in the MARAS procedure to remove aptamers with non-specific binding to unwanted biomolecules. Smart aptamers with predetermined affinity for C-reactive protein were isolated by window-MARAS. The specificity of aptamers was validated by using blind serum samples and compared with those using monoclonal antibody-based nephelometry analysis. The relative specificity of aptamers with or without primers was evaluated, revealing that the specificity of the aptamer with primer is similar to that of the aptamer without primer. By using a randomized library with primers, the simplicity and rapidity of MARAS in generating aptamers are preserved, and by implementing multiple negative selection, MARAS could efficiently select aptamers with high binding affinity and specificity for clinical applications.
机译：具有引物结合位点的适体对于SELEX（通过指数富集的配体的系统演化）过程是必需的。这种引物序列因其性质而导致非特异性结合，并提高了误报的发生率。因此，在没有引物或最小底漆的情况下使用适体，以缩短所选适体的长度，已成为一个有趣的研究主题。据报道，据报道，基于磁辅助快速适体选择（MARAS）技术的无引物适体选择方案以产生具有高亲和力和特异性的基底自由素。不对不利地，合适的适体的产率低，玛拉斯的优点劣化。在该研究中，在MARAS过程中使用多个负选择运行以除去具有非特异性结合到不需要的生物分子的适体。通过窗口玛拉斯分离具有预定亲和力的具有预定亲和力的智能适体。通过使用盲血清样品验证适体的特异性，与使用基于单克隆抗体的幼儿测定分析的人进行比较。具有或不具有引物适体的相对特异性进行评价，揭示了用引物适体的特异性类似于没有底漆的适体。通过使用具有引物的随机文库，保留了产生适体的玛拉斯的简单性和快速性，并且通过实施多个阴性选择，Maras可以有效地选择具有高结合亲和力和临床应用的特异性的适体。

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  • 来源
    《RSC Advances》 |2017年第68期|共10页
  • 作者

    Li Huan-Hao; Wen Chih-Yung; Hong Chin-Yih; Lai Ji-Ching;

  • 作者单位

    Hong Kong Polytech Univ Dept Mech Engn Hong Kong Hong Kong Peoples R China;

    Hong Kong Polytech Univ Dept Mech Engn Hong Kong Hong Kong Peoples R China;

    Natl Chung Hsing Univ Grad Inst Biomed Engn Taichung Taiwan;

    Natl Taiwan Normal Univ Inst Electroopt Sci &

    Technol Taipei Taiwan;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 化学;
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