Development of a sensitive micro-magnetic chemiluminescence enzyme immunoassay for the determination of carcinoembryonic antigen

Anal Bioanal Chem. 2007 Mar;387(6):1965-71. doi: 10.1007/s00216-006-0899-y. Epub 2006 Nov 18.

Abstract

A micro-magnetic chemiluminescence (CL) enzyme immunoassay with high sensitivity, selectivity, and reproducibility was developed for the determination of the tumor marker, carcinoembryonic antigen (CEA) in human serum. A sandwich scheme assay has been utilized with fluorescein isothiocyanate antibody (FITC)-labeled anti-CEA antibody and alkaline phosphate (ALP)-labeled anti-CEA antibody being used in the CL detection. The CL signal produced by the emission of photons from 4-methoxy-4-(3-phosphate-phenyl)-spiro-(1,2-dioxetane-3,2'-adamantane) (AMPPD) was directly proportional to the amount of analyte present in a sample solution. The influences of the reaction time of antigen with antibody, the reaction time of substrate with label, the dilution ratio of ALP-labeled anti-CEA antibody, the concentration of FITC-labeled anti-CEA antibody, and other relevant variables upon the CL signal were examined and optimized. The CL responses depended linearly on the CEA concentration over the range from 2 to 162 ng mL-1 in a logarithmic plot. Assay sensitivity as low as 0.69 ng mL-1 was achieved. A coefficient of variance of less than 13% was obtained for intra- and inter-assay precision. This method has been successfully applied to the analysis of CEA in human serum. According to the procedure based on spiked standards, the recoveries obtained were 80-110%. Comparison experiments were carried out with the commercially available CEA chemiluminescence immunoassay. Satisfactory results were obtained according to a paired t-test method (t value<tcritical at the 95% confidence level).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calibration
  • Carcinoembryonic Antigen / blood*
  • Carcinoembryonic Antigen / immunology
  • Chemical Phenomena
  • Chemistry, Physical
  • Humans
  • Immunoassay / methods*
  • Indicator Dilution Techniques
  • Luminescent Measurements / methods*
  • Magnetics*
  • Sensitivity and Specificity
  • Substrate Specificity
  • Time Factors

Substances

  • Carcinoembryonic Antigen