Guo Wenfei, Wang Haiyuan, Wang Zhaoyang, Wu Fandi, He Yao, Liu Yuan, Deng Yan, Bing Tao, Qiu Liping, Tan Weihong
Molecular Science and Biomedicine Laboratory (MBL), State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, Aptamer Engineering Center of Hunan Province, Hunan University, Changsha, Hunan, 410082, China; Zhejiang Cancer Hospital, Hangzhou Institute of Medicine (HIM), The Chinese Academy of Sciences, Hangzhou, Zhejiang, 310022, China.
Molecular Science and Biomedicine Laboratory (MBL), State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, Aptamer Engineering Center of Hunan Province, Hunan University, Changsha, Hunan, 410082, China.
Biosens Bioelectron. 2025 Mar 1;271:116996. doi: 10.1016/j.bios.2024.116996. Epub 2024 Nov 26.
Nicotinamide-adenine dinucleotide (NAD(H)) plays a critical role in cellular metabolism, and its accurate measurement is essential for elucidating biological mechanisms and disease progression. However, specific recognition probes and sensitive biosensors for NAD(H) remains a significant challenge. Here, we screen an aptamer (NAD3-1a) that exhibits specific binding to NAD(H) with micromolar affinity. By incorporating this aptamer with tetrahedral DNA nanostructure, we develop a highly selective and sensitive electrochemical biosensor for the detection of NAD(H). This biosensor enables precise detection of NAD(H) within a linear range of 10 ∼ 10 M, offering remarkable stability and reproducibility. Utilizing this biosensor, we observed significant variations in the NAD(H) levels between normal and tumor cells, as well as a notable reduction in NAD(H) in the skeletal muscle tissues of aged mice. These results highlight the potential of this aptamer-based biosensor to advance our understanding of metabolic variations in both health and disease contexts.
烟酰胺腺嘌呤二核苷酸（NAD(H)）在细胞代谢中起关键作用，其准确测量对于阐明生物学机制和疾病进展至关重要。然而，用于NAD(H)的特异性识别探针和灵敏生物传感器仍然是一项重大挑战。在此，我们筛选出一种适体（NAD3-1a），它能以微摩尔亲和力与NAD(H)特异性结合。通过将这种适体与四面体DNA纳米结构相结合，我们开发出一种用于检测NAD(H)的高选择性和灵敏的电化学生物传感器。该生物传感器能够在10~10 M的线性范围内精确检测NAD(H)，具有出色的稳定性和重现性。利用这种生物传感器，我们观察到正常细胞与肿瘤细胞之间NAD(H)水平存在显著差异，并且老年小鼠骨骼肌组织中的NAD(H)明显降低。这些结果凸显了这种基于适体的生物传感器在推进我们对健康和疾病背景下代谢变化理解方面的潜力。
