Development of an immunomagnetic bead-based time-resolved fluorescence immunoassay for rapid determination of levels of carcinoembryonic antigen in human serum.
A novel immunoassay for the determination of tumor markers in human serum was established by combining a time-resolved fluoroimmunoassay (TRFIA) and immunomagnetic separation. Based on a sandwich-type immunoassay format, analytes in samples were captured by magnetic beads coated with one monoclonal antibody and "sandwiched" by another monoclonal antibody labeled with europium chelates. The immunocomplex was separated and washed by exposure to a magnetic field and treatment with enhancement solution; fluorescence was then measured according to the number of europium ions dissociated. Levels of the model analyte, carcinoembryonic antigen (CEA), were determined in a linear range (1-1000 ng mL(-1)) with a limit of detection of 0.5 ng mL(-1) under optimal conditions. The reproducibility, recovery, and specificity of the immunoassay were demonstrated to be acceptable. To evaluate this novel assay for clinical applications, 239 serum samples were evaluated. Compared with the conventional TRFIA and chemiluminescence immunoassay (CLIA), the correlation coefficients of the developed immunoassay were 0.985 and 0.975, respectively. These results showed good correlation and confirmed that our method is feasible and could be used for the clinical determination of CEA (or other tumor antigens) in human serum.
Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou 510515, PR China.
Jing-Yuan
JY
Hou
Tian-Cai
TC
Liu
Guan-Feng
GF
Lin
Zhi-Xiong
ZX
Li
Li-Ping
LP
Zou
Ming
M
Li
Ying-Song
YS
Wu
D014408:Biomarkers, Tumor / Q000097:blood*; D002272:Carcinoembryonic Antigen / Q000097:blood*; D015200:Fluoroimmunoassay / Q000379:methods*; D006801:Humans; D018189:Immunomagnetic Separation; D008954:Models, Biological; D015203:Reproducibility of Results; D013997:Time Factors
D014408:Biomarkers, Tumor; D002272:Carcinoembryonic Antigen