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Evaluation of different strategies for magnetic particle functionalization with DNA aptamers

机译：使用DNA适体评估磁性粒子功能化的不同策略

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The optimal bio-functionalization of magnetic particles is essential for developing magnetic particle-based bioassays. Whereas functionalization with antibodies is generally well established, immobilization of DNA probes, such as aptamers, is not yet fully explored. In this work, four different types of commercially available magnetic particles, coated with streptavidin, maleimide or carboxyl groups, were evaluated for their surface coverage with aptamer bioreceptors, efficiency in capturing target protein and non-specific protein adsorption on their surface. A recently developed aptamer against the peanut allergen, Ara h 1 protein, was used as a model system. Conjugation of biotinylated Ara h 1 aptamer to the streptavidin particles led to the highest surface coverage, whereas the coverage of maleimide particles was 25% lower. Carboxylated particles appeared to be inadequate for DNA functionalization. Streptavidin particles also showed the greatest target capturing efficiency, comparable to the one of particles functionalized with anti-Ara h 1 antibody. The performance of streptavidin particles was additionally tested in a sandwich assay with the aptamer as a capture receptor on the particle surface. While the limit of detection obtained was comparable to the same assay system with antibody as capture receptor, it was superior to previously reported values using the same aptamer in similar assay schemes with different detection platforms. These results point to the promising application of the Ara h 1 aptamer-functionalized particles in bioassay development. (C) 2016 Elsevier B.V. All rights reserved.
机译：磁性粒子的最佳生物功能化对于开发基于磁性粒子的生物测定至关重要。尽管通常已经很好地确定了用抗体进行的功能化，但尚未充分探索DNA探针（如适体）的固定化。在这项工作中，对涂有链霉亲和素，马来酰亚胺或羧基的四种不同类型的可商购磁性粒子的适体生物受体表面覆盖率，捕获靶蛋白的效率和非特异性蛋白在其表面的吸附进行了评估。最近开发的针对花生过敏原的适体Ara h 1蛋白被用作模型系统。生物素化的Ara h 1适体与链霉亲和素颗粒的缀合导致最高的表面覆盖率，而马来酰亚胺颗粒的覆盖率低25％。羧基化的颗粒似乎不足以实现DNA功能化。与抗Ara h 1抗体功能化的颗粒之一相比，链霉亲和素颗粒还显示出最大的靶标捕获效率。链霉亲和素颗粒的性能另外在夹心测定中用适体作为颗粒表面上的捕获受体进行了测试。尽管获得的检测限可与使用抗体作为捕获受体的相同检测系统相媲美，但它优于先前报道的值，在具有不同检测平台的相似检测方案中使用相同的适体。这些结果表明Ara h 1适体功能化的粒子在生物测定开发中的有希望的应用。 （C）2016 Elsevier B.V.保留所有权利。

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