Improving CEA detection Sensitivity: Carboxyfluorescein-Loaded liposomes in aptamer sandwich assay

This study aimed to develop an effective method for highly sensitive detection of carcinoembryonic antigen (CEA), a tumor biomarker, using an aptasensor sandwich assay based on a liposome loaded with carboxyfluorescein. In this approach, liposomes loaded with carboxyfluorescein (CF) were anchored with a cholesterol-labeled anti-CEA aptamer (Apt1-Lip) using a post-insertion technique. Furthermore, the successful insertion of the aptamer into the liposome outer layer was confirmed through UV absorption, PAGE electrophoresis, dynamic light scattering (DLS), and zeta potential measurements. Additionally, a secondary aptamer was conjugated with magnetic beads via biotin-streptavidin interactions and used as a capture probe (MBs/Apt2). Aptamer-decorated liposomes successfully bound to the CEA antigen in the presence of the target, leading to the formation of a sandwich assay when combined with MBs/Apt2. The specific binding signal was further increased by dissolving the lipid nanospheres with Triton X-100. This liposome-based aptasensor demonstrated a detection limit of 35.48 pg/mL over a wide linear range from 0.1 ng/mL to 100 ng/mL, with high specificity and sensitivity at CEA. Additionally, the proposed approach exhibited notable promise for on-site testing in early clinical diagnosis, as it showed excellent selectivity, satisfactory reproducibility, and good recovery in CEA detection from human serum samples.