Immunological assay for carbohydrate antigen 19-9 using an electrochemical immunosensor and antigen immobilization in titania sol-gel matrix

J Immunol Methods. 2003 Dec;283(1-2):67-75. doi: 10.1016/j.jim.2003.08.014.

Abstract

We describe a novel electrochemical immunosensor for carbohydrate antigen 19-9 (CA19-9) based on the immobilization of CA19-9 with titania sol-gel on a graphite electrode (GE) by vapor deposition. The CA19-9 membrane was characterized using scanning electron microscopy and proved to be chemically clean, porous and homogeneous. The incubation of the immunosensor in a solution containing horseradish peroxidase (HRP)-labeled CA19-9 antibody led to the binding of HRP-labeled antibody with the immobilized antigen. The immobilized HRP catalyzed the oxidation of catechol by H(2)O(2) and this provided a competitive method for the measurement of serum CA19-9. The response current decreased with increasing CA19-9 concentration in the incubation solution. The effects of pH, amount of HRP-labeled antibody, incubation time and temperature were explored to provide optimum analytical performance. Under optimal conditions, the current decrease of the immunosensor was proportional to CA19-9 concentrations in the range of 3-20 U/ml with a detection limit of 2.68 U/ml at a current decrease of 10%. The detection of CA19-9 in two serum samples obtained from clinically diagnosed patients with pancreatic carcinoma showed acceptable accuracy. The proposed immunosensor provides a new promising tool for the clinical immunoassay of CA19-9.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques / methods*
  • CA-19-9 Antigen / blood*
  • Electrochemistry
  • Gels
  • Immunoassay / methods*
  • Microscopy, Electron, Scanning
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Titanium

Substances

  • CA-19-9 Antigen
  • Gels
  • titanium dioxide
  • Titanium