Wu Xingjie, Xiong Shasha, Tao Ling, Huang Jian, Shen Xiangchun
State Key Laboratory of Functions and Applications of Medicinal Plants, School of Pharmaceutical Sciences, Guizhou Medical University, Guiyang City and Guian New District, No.6 Ankang Avenue, Guizhou, 561113, China.
The High Efficacy Application of Natural Medicinal Resources Engineering Center of Guizhou Province, School of Pharmaceutical Sciences, Guizhou Medical University, Guiyang City and Guian New District, No.6 Ankang Avenue, Guizhou, 561113, China.
Mikrochim Acta. 2024 Dec 21;192(1):21. doi: 10.1007/s00604-024-06885-1.
A novel glycoprotein assay was developed by integrating the hairpin aptamer (H-APT)-mediated glycoprotein recognition and the reactive oxygen species-sensitive microcapsule (ROS-MC)-induced signal amplification. The analyzing process begins with the transfer of the target glycoprotein to a chlorin e6 (Ce6)-labeled DNA sequence via H-APT-mediated DNA displacement. Subsequently, the Ce6-labeled DNA was used to induce the disassembly of fluorophore-loaded ROS-MC under 650-nm light irradiation. Leveraging the rapid release of the fluorophore and the high loading capacity of the MC, this glycoprotein assay is capable of quantifying glycoprotein content in native biofluids within 2.5 h, achieving a detection limit of 0.034 ng/mL. We applied this assay to determine the glycoprotein composition in plasma samples of colorectal cancer patients, revealing a significant increase in glycoprotein content for those with a poor prognosis. In summary, we have developed an innovative method for glycoprotein determination that shows potential for clinical translation.
通过整合发夹适体(H-APT)介导的糖蛋白识别和活性氧敏感微胶囊(ROS-MC)诱导的信号放大,开发了一种新型糖蛋白检测方法。分析过程始于通过H-APT介导的DNA置换将目标糖蛋白转移到叶绿素e6(Ce6)标记的DNA序列上。随后,在650纳米光照射下,用Ce6标记的DNA诱导负载荧光团的ROS-MC解体。利用荧光团的快速释放和微胶囊的高负载能力,这种糖蛋白检测方法能够在2.5小时内定量天然生物流体中的糖蛋白含量,检测限达到0.034纳克/毫升。我们应用该检测方法测定结直肠癌患者血浆样本中的糖蛋白组成,发现预后不良患者的糖蛋白含量显著增加。总之,我们开发了一种创新的糖蛋白测定方法,具有临床转化潜力。