Choi Ji Eun, Yun Hanool, Jeong Hee-Jin
Department of Pathology, Chungnam National University Sejong Hospital, Sejong 30099, Republic of Korea.
Department of Biological and Chemical Engineering, Hongik University, Sejong 30016, Republic of Korea.
Curr Issues Mol Biol. 2025 Jan 17;47(1):62. doi: 10.3390/cimb47010062.
The development of accurate and high-throughput tools for cancer biomarker detection is crucial for the diagnosis, monitoring, and treatment of diseases. In this study, we developed a simple and rapid fluorescence-linked immunosorbent assay (FLISA) using fluorescent dye-conjugated antibody fragments against programmed cell death ligand 1 (PDL1) and human epithelial growth factor receptor 2 (HER2). We optimized key steps in the FLISA process, including antigen immobilization, blocking, and antibody reaction, reading the assay time to 3 h-significantly faster compared to the 23 h duration of usual FLISA. The limit of detection for the rapid FLISA in detecting PDL1 was lower than that of FLISA, and the detection of HER2 was similar between the two methods, indicating that the rapid FLISA provides a fast and accurate approach for detecting PDL1 and HER2. This robust platform can be readily adapted for various fluoroimmunoassays targeting other antigens of interest.
开发用于癌症生物标志物检测的准确且高通量的工具对于疾病的诊断、监测和治疗至关重要。在本研究中,我们使用针对程序性细胞死亡配体1(PDL1)和人表皮生长因子受体2(HER2)的荧光染料偶联抗体片段,开发了一种简单快速的荧光连接免疫吸附测定法(FLISA)。我们优化了FLISA过程中的关键步骤,包括抗原固定、封闭和抗体反应,将检测时间缩短至3小时,与常规FLISA的23小时相比明显更快。快速FLISA检测PDL1的检测限低于FLISA,两种方法检测HER2的结果相似,这表明快速FLISA为检测PDL1和HER2提供了一种快速准确的方法。这个强大的平台可以很容易地适用于针对其他感兴趣抗原的各种荧光免疫测定。