Zhang Yarong, Ma Hongyan, Gao Zhenyao, Li Miao, Yang Fan, Sun Lingbo, Zhang Yuecheng
Key Laboratory of Analytical Technology and Detection of Yan'an, College of Chemistry and Chemical Engineering, Yan'an University, Yan'an 716000, China.
Medical College of Yan'an University, Yan'an University, Yan'an 716000, China.
Biomolecules. 2025 Sep 3;15(9):1275. doi: 10.3390/biom15091275.
Apurinic/apyrimidinic endonuclease 1 (APE1) selectively cleaves the apurinic/apyrimidinic site (AP site) in DNA, playing a critical role in base excision repair and genomic stability maintenance. Aberrant APE1 expression has been linked to numerous diseases, including cardiovascular disorders, neurological conditions, and various cancers. However, existing methods for detecting trace levels of APE1 remain suboptimal for certain applications. To address this limitation, we developed an innovative biosensing platform for ultrasensitive APE1 detection by integrating APE1-specific site recognition with hybridization chain reaction (HCR)-based signal amplification, enabling enzyme- and label-free bioassays. In this system, APE1 recognizes and cleaves the AP site-containing hairpin (HP) probe, releasing a single-stranded HCR initiator that triggers cascaded HCR amplification. Owing to the high efficiency of HCR, this method achieves ultrahigh sensitivity, with a calculated detection limit of 1.0 × 10 U/mL. Furthermore, the biosensor demonstrates robust performance in cell lysates and is applicable for screening and evaluating APE1 inhibitors.
脱嘌呤/脱嘧啶内切酶1（APE1）选择性切割DNA中的脱嘌呤/脱嘧啶位点（AP位点），在碱基切除修复和基因组稳定性维持中发挥关键作用。APE1表达异常与多种疾病相关，包括心血管疾病、神经疾病和各种癌症。然而，现有的检测痕量APE1的方法在某些应用中仍不尽人意。为解决这一局限性，我们通过将APE1特异性位点识别与基于杂交链式反应（HCR）的信号放大相结合，开发了一种用于超灵敏检测APE1的创新生物传感平台，实现了无酶和无标记的生物检测。在该系统中，APE1识别并切割含AP位点的发夹（HP）探针，释放出单链HCR引发剂，触发级联HCR扩增。由于HCR的高效性，该方法实现了超高灵敏度，计算得到的检测限为1.0×10 U/mL。此外，该生物传感器在细胞裂解物中表现出强大的性能，适用于APE1抑制剂的筛选和评估。
