Establishment of a Rapid and Convenient Fluoroimmunoassay Platform Using Antibodies Against PDL1 and HER2

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.