4.7 Article

Development of an immunomagnetic bead-based time-resolved fluorescence immunoassay for rapid determination of levels of carcinoembryonic antigen in human serum

Journal

ANALYTICA CHIMICA ACTA
Volume 734, Issue -, Pages 93-98

Publisher

ELSEVIER SCIENCE BV
DOI: 10.1016/j.aca.2012.04.044

Keywords

Magnetic beads; Time-resolved fluoroimmunoassay; Carcinoembryonic antigen

Funding

  1. National Natural Science Foundation of China [30901382]
  2. New Teacher for Doctoral Fund of Ministry of Education of China [2009443312008]
  3. Special Funds for Colleges and Universities Talents by Guangdong Province

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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. (c) 2012 Elsevier B.V. All rights reserved.

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