Zhang Liang, Zhao Pinnan, Liu Yujun, Shi Ning, Zhou Yangyihua, Peng Shangde, Sun Tieqiang, Zhang Min, Wu Yahui, Yang Xuechen, Wen Yan, Shi Gang, Gao Xiang, Luo Longlong
College of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang, 212003, China; State Key Laboratory of Toxicology and Medical Countermeasures, Beijing Institute of Pharmacology and Toxicology, Beijing, 100850, China.
State Key Laboratory of Toxicology and Medical Countermeasures, Beijing Institute of Pharmacology and Toxicology, Beijing, 100850, China.
Talanta. 2025 Apr 1;285:127301. doi: 10.1016/j.talanta.2024.127301. Epub 2024 Dec 4.
Tumor necrosis factor alpha (TNF-α) is a key cytokine in inflammation and immune responses, making its rapid and accurate detection essential for disease diagnosis and management. In this study, we developed a highly sensitive chemiluminescence immunoassay (CLIA) using antibody-coated magnetic particles (Ab-MPs-CLIA) for TNF-α detection. From nine candidate antibodies, we identified an optimal pair through epitope competition and affinity assessments, significantly improving assay performance. The Ab-MPs-CLIA achieved a detection limit of 0.25 pg/mL, 6.8 times more sensitive than Siemens commercial kits, with a broad linear range of 9.2-1077 pg/mL. The method demonstrated excellent stability, both under accelerated conditions at 37 °C for 7 days and long-term storage at 4 °C for 12 months. It showed no cross-reactivity with common interfering substances in human serum, ensuring high specificity. Notably, the entire process, from sample preparation to result, takes just 25 min, compared to 3-4 h for both ELISA and RIA, and CLIA typically offers 10-100 times higher sensitivity than these methods. These advantages make the Ab-MPs-CLIA an ideal option for clinical laboratories, providing superior sensitivity, specificity, broader dynamic range, and greater operational efficiency than existing TNF-α detection technologies.
肿瘤坏死因子α（TNF-α）是炎症和免疫反应中的关键细胞因子，因此对其进行快速准确的检测对于疾病的诊断和管理至关重要。在本研究中，我们开发了一种使用抗体包被磁颗粒的高灵敏度化学发光免疫分析法（Ab-MPs-CLIA）用于检测TNF-α。我们从九种候选抗体中，通过表位竞争和亲和力评估确定了一对最佳抗体，显著提高了检测性能。Ab-MPs-CLIA的检测限为0.25 pg/mL，比西门子商业试剂盒灵敏6.8倍，线性范围宽达9.2 - 1077 pg/mL。该方法在37°C加速条件下放置7天以及在4°C长期保存12个月时均表现出优异的稳定性。它与人血清中的常见干扰物质无交叉反应，确保了高特异性。值得注意的是，从样品制备到得出结果的整个过程仅需25分钟，而酶联免疫吸附测定法（ELISA）和放射免疫测定法（RIA）都需要3 - 4小时，并且化学发光免疫分析法（CLIA）的灵敏度通常比这些方法高10 - 100倍。这些优势使Ab-MPs-CLIA成为临床实验室的理想选择，与现有的TNF-α检测技术相比，它具有更高的灵敏度、特异性、更宽的动态范围和更高的操作效率。
