Kumaragurubaran Namasivayam, Yun Hou Po, Zinovicius Antanas, Huang Sheng-Tung, Arul Ponnusamy, Lin Hsin-Yi, Min-Shin Ou, Morkvenaite-Vilkonciene Inga
Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei, 106, Taiwan, ROC; Institute of Biochemical and Biomedical Engineering, National Taipei University of Technology, Taipei, 106, Taiwan, ROC.
Department of Nanotechnology, State Research Institute Centre for Physical Sciences and Technology (FTMC), Sauletekio av. 3, 10257, Vilnius, Lithuania.
Biosens Bioelectron. 2025 Sep 1;283:117538. doi: 10.1016/j.bios.2025.117538. Epub 2025 May 2.
Dipeptidyl peptidase-IV (DPP-IV) is a circulating blood biomarker that diagnose pancreatic and thyroid cancers, as well as type 2 diabetes. Although current DPP-IV detection methods show promise, real-time detection in whole blood is limited, as blood samples require tedious pre-treatment. To overcome these limitations, a DPP-IV targeted electrochemical substrate, DPPLPOH (DiPeptidyl Peptidase Latent Probe-OH (with Hydroxyl group)), was designed. When coupled with an electrochemical analytical method, this substrate enabled direct and convenient detection of DPP-IV in complex biofluids, including whole blood samples. In these assays, DPP-IV selectively hydrolyzed DPPLPOH, which underwent a self-immolative reaction to generate a masked electrochemically sensitive amino ferrocene reporter (AFOH). This electrochemical analytical tool demonstrated excellent sensing performance, characterized by exceptional enzyme binding properties. DPPLPOH showed excellent sensitivity and selectivity, with a detection limit of 0.021 ng/mL and a broad linear detection range of 0.1-100 ng/mL. The probe was specific to DPP-IV without interference from other electroactive species, enzymes, or hydrolases. Furthermore, DPPLPOH enabled real-time monitoring of DPP-IV activity on tumor cell surfaces and direct tracking of DPP-IV concentration in whole blood without a tedious separation process. This method may be a valuable tool in the early detection of pancreatic and thyroid cancers and in post-treatment surveillance.
二肽基肽酶-IV（DPP-IV）是一种循环血液生物标志物，可用于诊断胰腺癌、甲状腺癌以及2型糖尿病。尽管目前的DPP-IV检测方法显示出了前景，但全血中的实时检测受到限制，因为血液样本需要繁琐的预处理。为了克服这些限制，设计了一种靶向DPP-IV的电化学底物DPPLPOH（二肽基肽酶潜伏探针-OH（含羟基））。当与电化学分析方法结合使用时，这种底物能够直接、方便地检测复杂生物流体（包括全血样本）中的DPP-IV。在这些检测中，DPP-IV选择性地水解DPPLPOH，后者发生自牺牲反应，生成一个被掩蔽的电化学敏感氨基二茂铁报告分子（AFOH）。这种电化学分析工具表现出优异的传感性能，其特点是具有出色的酶结合特性。DPPLPOH显示出优异的灵敏度和选择性，检测限为0.021 ng/mL，线性检测范围宽，为0.1 - 100 ng/mL。该探针对DPP-IV具有特异性，不受其他电活性物质、酶或水解酶的干扰。此外，DPPLPOH能够实时监测肿瘤细胞表面的DPP-IV活性，并直接追踪全血中DPP-IV的浓度，无需繁琐的分离过程。这种方法可能是早期检测胰腺癌和甲状腺癌以及治疗后监测的有价值工具。
