Zhou Na, Zeng Yu, Huang Qiuwen, Tan Rui, Chen Yushi, Zhong Liu, Luo Xiaojun, Deng Xiaojun
School of Science, Xihua University, Chengdu, 610039, PR China.
School of Science, Xihua University, Chengdu, 610039, PR China.
Anal Chim Acta. 2025 Sep 15;1367:344307. doi: 10.1016/j.aca.2025.344307. Epub 2025 Jun 6.
BACKGROUND: The misuse of recombinant human erythropoietin-alpha (rHuEPO-α) for performance enhancement in sports has been a persistent issue. The World Anti-Doping Agency has emphasized the need for reliable detection methods to identify the illicit use of rHuEPO-α, thereby upholding the integrity of sports competitions. Most screening strategies rely on immunological methods for detection. However, these methods often depend on antibodies with limited selectivity, leading to a higher risk of false-positive results. Consequently, there is a pressing need to develop alternative and innovative recognition modules for the precise detection of rHuEPO-α. Aptamers, often termed "chemical antibodies," are well-suited to address this challenge. RESULTS: The Raman signaling molecule 4-aminothiophene (4-ATP) was initially embedded between gold-silver core-shell nanostars (Au@Ag NSs). The rHuEPO-α aptamer was then conjugated to the Ag shell via an Ag-S covalent bond to assemble the Surface-Enhanced Raman Scattering (SERS) probe. Subsequently, the capture DNA strand was connected to the magnetic covalent organic framework (FeO@COF@Au) via "Au-S" bonds. The complementary pairing of the aptamer on the SERS probe with the captured DNA enabled the assembly of the magnetic covalent organic framework and the SERS probe. The specific recognition of rHuEPO-α by the aptamer triggered the separation of the SERS probe from the FeO@COF@Au magnetic covalent organic framework, resulting in a decrease in the Raman signal. The detection limit for rHuEPO-α concentration was ultimately determined to be as low as 0.856 pg/mL, exhibiting an inverse linear relationship with the Raman signal. SIGNIFICANCE: The aptasensor utilizes its magnetic separation effect to enhance the enrichment and purification of Raman signals, thereby improving detection sensitivity and achieving a lower detection limit. This provides an experimental basis for the construction of SERS aptasensors and the detection of rHuEPO-α. Moreover, the sensor also has the potential to serve as an effective monitoring tool for other trace substances.
背景：重组人促红细胞生成素-α（rHuEPO-α）在体育赛事中被滥用以提高成绩一直是个持续存在的问题。世界反兴奋剂机构强调需要可靠的检测方法来识别rHuEPO-α的非法使用，从而维护体育比赛的公正性。大多数筛查策略依赖免疫方法进行检测。然而，这些方法通常依赖选择性有限的抗体，导致假阳性结果的风险更高。因此，迫切需要开发用于精确检测rHuEPO-α的替代和创新识别模块。适配体，通常被称为“化学抗体”，非常适合应对这一挑战。 结果：拉曼信号分子4-氨基噻吩（4-ATP）最初嵌入金银核壳纳米星（Au@Ag NSs）之间。然后，rHuEPO-α适配体通过Ag-S共价键与Ag壳共轭，以组装表面增强拉曼散射（SERS）探针。随后，捕获DNA链通过“Au-S”键连接到磁性共价有机框架（FeO@COF@Au）。SERS探针上的适配体与捕获DNA的互补配对实现了磁性共价有机框架和SERS探针的组装。适配体对rHuEPO-α的特异性识别触发了SERS探针与FeO@COF@Au磁性共价有机框架的分离，导致拉曼信号降低。rHuEPO-α浓度的检测限最终确定低至0.856 pg/mL，与拉曼信号呈反比线性关系。 意义：该适配体传感器利用其磁分离效应增强拉曼信号的富集和纯化，从而提高检测灵敏度并实现更低的检测限。这为构建SERS适配体传感器和检测rHuEPO-α提供了实验依据。此外，该传感器还具有作为其他痕量物质有效监测工具的潜力。
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