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一种基于适体的基因场效应晶体管传感器,用于特异性定量基因治疗型人类5型腺病毒。

An Aptamer-Based gFET-Sensor for Specific Quantification of Gene Therapeutic Human Adenovirus Type 5.

作者信息

Li Runliu, Kissmann Ann-Kathrin, Xing Hu, Hasler Roger, Kleber Christoph, Knoll Wolfgang, Schmietendorf Hannes, Engler Tatjana, Krutzke Lea, Kochanek Stefan, Rosenau Frank

机构信息

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

Faculty of Medicine and Dentistry, Danube Private University, Steiner Landstraße 124, 3500 Krems an der Donau, Austria.

出版信息

Biosensors (Basel). 2025 Sep 14;15(9):605. doi: 10.3390/bios15090605.

Abstract

The combination of rGO-FETs (reduced Graphene Oxide Field-Effect Transistors) and DNA-oligonucleotide aptamers to sense analytes has been shown to be a promising technological approach, achieving high sensitivity and selectivity. With human adenovirus type 5 (HAdV-5) particles as the target, we here demonstrate the application of the aptamer/FET combination for detection of this medically and biotechnologically relevant viral vector. A focused anti-HAdV-5 aptamer library was evolved in a nine-round SELEX process, allowing for the specific fluorescent labeling of HAdV-5 and related subtypes. Moreover, this library was already sufficient to serve as the binding entity on a gFET sensor for sensitive quantification of the virus particles. Adenoviruses have been widely used as gene delivery vectors for gene therapy and genetic vaccination. The use of adenoviral vectors within the vaccination campaign against COVID-19 emphasized the need for robust biotechnological production processes, which additionally require sensitive product formation monitoring. We believe that these type of gFET-based aptasensors can serve as the technological monitoring basis in virus production processes in the near future.

摘要

还原氧化石墨烯场效应晶体管(rGO - FETs)与DNA寡核苷酸适配体相结合用于传感分析物,已被证明是一种很有前景的技术方法,能够实现高灵敏度和高选择性。以5型人类腺病毒(HAdV - 5)颗粒为目标,我们在此展示了适配体/FET组合在检测这种医学和生物技术相关病毒载体中的应用。通过九轮SELEX过程筛选出了一个聚焦的抗HAdV - 5适配体文库,可实现对HAdV - 5及相关亚型的特异性荧光标记。此外,该文库已足以作为gFET传感器上的结合实体,用于灵敏定量病毒颗粒。腺病毒已被广泛用作基因治疗和基因疫苗接种的基因递送载体。在抗击新冠疫情的疫苗接种活动中使用腺病毒载体凸显了对稳健生物技术生产工艺的需求,这还额外需要对产品形成进行灵敏监测。我们相信,这类基于gFET的适配体传感器在不久的将来可作为病毒生产过程中的技术监测基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/08b9/12467633/ecf30483edd7/biosensors-15-00605-g001.jpg

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