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Highly sensitive and specific graphene oxide-based FRET aptasensor for quantitative detection of human soluble growth stimulating gene protein 2
Talanta ( IF 5.6 ) Pub Date : 2024-01-09 , DOI: 10.1016/j.talanta.2024.125629 Zhikun Zeng 1 , Wenfeng Li 2 , Jixuan Zhang 2 , Zijian Hu 3 , Junyi Wu 2 , Guangming Ye 1 , Yi Luo 1
Talanta ( IF 5.6 ) Pub Date : 2024-01-09 , DOI: 10.1016/j.talanta.2024.125629 Zhikun Zeng 1 , Wenfeng Li 2 , Jixuan Zhang 2 , Zijian Hu 3 , Junyi Wu 2 , Guangming Ye 1 , Yi Luo 1
Affiliation
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Soluble growth stimulation expressed gene 2 (sST2) is a new generation biomarker in the diagnosis and prognosis of heart failure (HF). Here, the sST2-specific aptamers were selected from a random ssDNA library with the full length of 88 nucleotides (nt) via target-immobilized magnetic beads (MB)-based systematic evolution of ligands by exponential enrichment (SELEX) technology. After eight rounds of selection, six aptamers with the most enrichment were selected. Among, the aptamer L1 showed the high-affinity binding to sST2 with the lowest Kd value (77.3 ± 0.05 nM), which was chosen as the optimal aptamer for further molecular docking. Then, the aptamer L1 was used to construct a graphene oxide (GO) - based fluorescence resonance energy transfer (FRET) biosensor for sST2, which exhibits a linear detection range of 0.1–100 μg/ml and a detection limit of 3.7 ng/ml. The aptasensor was applied to detect sST2 in real samples, with a good correlation and agreement with the traditional enzyme-linked immunosorbent assay (ELISA) when quantitative analyzing the sST2 concentration in serum samples from HF patients. The results show that not only an efficient strategy for screening the practicable aptamer, but also a rapid and sensitive detection platform for sST2 were established.
中文翻译：
高灵敏、特异的基于氧化石墨烯的 FRET 适体传感器用于定量检测人类可溶性生长刺激基因蛋白 2
可溶性生长刺激表达基因2（sST2）是心力衰竭（HF）诊断和预后的新一代生物标志物。这里，通过基于目标固定磁珠（MB）的指数富集配体系统进化（SELEX）技术，从全长88个核苷酸（nt）的随机ssDNA库中选择sST2特异性适体。经过八轮筛选，选出富集度最高的 6 个适配体。其中，适配体L1与sST2具有高亲和力结合，Kd值最低（77.3±0.05 nM），被选为进一步分子对接的最佳适配体。然后，利用适配体L1构建了基于氧化石墨烯（GO）的sST2荧光共振能量转移（FRET）生物传感器，其线性检测范围为0.1-100 μg/ml，检测限为3.7 ng/ml 。应用适配体传感器检测真实样本中的sST2，在定量分析心力衰竭患者血清样本中的sST2浓度时，与传统酶联免疫吸附试验（ELISA）具有良好的相关性和一致性。结果表明，不仅建立了筛选实用适配体的有效策略，而且建立了快速、灵敏的sST2检测平台。
更新日期：2024-01-09
中文翻译：
高灵敏、特异的基于氧化石墨烯的 FRET 适体传感器用于定量检测人类可溶性生长刺激基因蛋白 2
可溶性生长刺激表达基因2（sST2）是心力衰竭（HF）诊断和预后的新一代生物标志物。这里，通过基于目标固定磁珠（MB）的指数富集配体系统进化（SELEX）技术，从全长88个核苷酸（nt）的随机ssDNA库中选择sST2特异性适体。经过八轮筛选，选出富集度最高的 6 个适配体。其中，适配体L1与sST2具有高亲和力结合，Kd值最低（77.3±0.05 nM），被选为进一步分子对接的最佳适配体。然后，利用适配体L1构建了基于氧化石墨烯（GO）的sST2荧光共振能量转移（FRET）生物传感器，其线性检测范围为0.1-100 μg/ml，检测限为3.7 ng/ml 。应用适配体传感器检测真实样本中的sST2，在定量分析心力衰竭患者血清样本中的sST2浓度时，与传统酶联免疫吸附试验（ELISA）具有良好的相关性和一致性。结果表明，不仅建立了筛选实用适配体的有效策略，而且建立了快速、灵敏的sST2检测平台。
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