High-affinity ssDNA aptamer and chemiluminescent aptasensor for TIMP-1 detection in human serum

作者：Qin Wang, Yanli Li, Lige Yao, Huiqin Li, Liuyan Zhang, Yingjie Wang, Jiayin Li, Tian Chen, Kun Chai, Junli Gao, Junshun Gao, Li Su, Xueming Li · 发表于：Analytical Sciences · 年份：2025 · DOI：10.1007/s44211-024-00673-w · 被引用次数：4 · 研究领域：Advanced biosensing and bioanalysis techniques、Protease and Inhibitor Mechanisms、Cell Adhesion Molecules Research

Abstract TIMP-1 (Tissue Inhibitor of Metalloproteinases-1) is a protein involved in regulating extracellular matrix (ECM) degradation. It is recognized as a significant biomarker for cancer diagnosis. This study aimed to develop and characterize a single-stranded DNA (ssDNA) aptamer targeting human TIMP-1 protein with high affinity and specificity. A magnetic beads-based SELEX process combined with qPCR was used to select aptamers over seven rounds. The enriched ssDNA library was analyzed using high-throughput sequencing to identify candidate sequences, and these sequences were characterized using surface plasmon resonance (SPR) and binding assays to evaluate their affinity and specificity. The selected ssDNA aptamer demonstrated a dissociation equilibrium constant ( K D ) of 0.41 nM and a very slow off-rate, enabling effective capture of TIMP-1 in serum samples. Furthermore, a chemiluminescent aptasensor was developed for TIMP-1 detection, which exhibited high specificity and a broad linear detection range from 1 to 500 ng/mL in human serum. The developed ssDNA aptamer targeting TIMP-1 shows high affinity and specificity, and the chemiluminescent aptasensor demonstrates promising potential for clinical diagnosis of TIMP-1 levels in human serum. Graphical abstract