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FluCell-SELEX 适体作为用于诊断与健康相关肠道细菌的特异性结合分子。

FluCell-SELEX Aptamers as Specific Binding Molecules for Diagnostics of the Health Relevant Gut Bacterium .

机构信息

Institute for Pharmaceutical Biotechnology, Ulm University, Albert-Einstein-Allee 11, 89081 Ulm, Germany.

Max Planck Institute for Polymer Research Mainz, Ackermannweg 10, 55128 Mainz, Germany.

出版信息

Int J Mol Sci. 2021 Sep 27;22(19):10425. doi: 10.3390/ijms221910425.

Abstract

Based on their unique properties, oligonucleotide aptamers have been named a gift of biological chemistry to life science. We report the development of DNA aptamers as the first high-affinity binding molecules available for fast and rapid labeling of the human gut bacterium with a certain impact on Alzheimer´s disease. Fast and reliable analyses of the composition of microbiomes is an emerging field in microbiology. We describe the molecular evolution and biochemical characterization of a specific aptamer library by a FluCell-SELEX and the characterization of specific molecules from the library by bioinformatics. The aptamer AKK13.1 exerted universal applicability in different analysis techniques in modern microbiology, including fluorimetry, confocal laser scanning microscopy and flow cytometry. It was also functional as a specific binding entity hybridized to anchor primers chemically coupled via acrydite-modification to the surface of a polyacrylamide-hydrogel, which can be prototypically used for the construction of affinity surfaces in sensor chips. Together, the performance and methodological flexibility of the aptamers presented here may open new routes not only to develop novel -specific assays for clinical microbiology and the analyses of human stool samples but may also be an excellent starting point for the construction of novel electronic biosensors.

摘要

基于它们独特的性质,寡核苷酸适体被称为生物化学对生命科学的馈赠。我们报告了 DNA 适体的开发,作为第一个可用于快速标记人类肠道细菌的高亲和力结合分子,对阿尔茨海默病有一定影响。快速可靠地分析微生物组的组成是微生物学中一个新兴的领域。我们通过 FluCell-SELEX 描述了一个特定适体文库的分子进化和生化特性,并通过生物信息学对文库中的特定分子进行了表征。适体 AKK13.1 在现代微生物学的不同分析技术中具有普遍适用性,包括荧光计、共聚焦激光扫描显微镜和流式细胞术。它还可以作为一种特异性结合实体,与通过丙烯酰胺修饰化学偶联到聚丙烯酰胺水凝胶表面的锚定引物杂交,这可以作为在传感器芯片上构建亲和表面的原型。总之,这里提出的适体的性能和方法学灵活性不仅可能为临床微生物学和人类粪便样本的分析开发新的特定检测方法开辟新途径,而且可能也是构建新型电子生物传感器的绝佳起点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d06f/8509069/ffe5da7a853e/ijms-22-10425-g001.jpg

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