An amperometric immunosensor based on a conducting immunocomposite electrode for the determination of Schistosoma japonicum antigen.

A renewable amperometric immunosensor based on a graphite-paraffin-Schistosoma japonicum antibody (SjAb) biocomposite electrode has been prepared for the detection of Schistosoma japonicum antigen (SjAg). Competitive ELISA was employed involving HRP-SjAg as a tracer and 3,3',5,5'-tetramethylbenzidine (TMB) as a substrate. The product of an enzyme catalytic reaction was detected at +0.1 V (vs. Ag/AgCl reference electrode) for measuring the amount of HRP-labeled SjAg binding to the electrode surface. The assay conditions were optimized, including the amount of SjAb loading in the electrode and HRP-SjAg in the incubation solution, the pH of the measuring solution and the incubation time. The measuring range was 0.5-30 microg/ml under the optimum conditions. Rabbit serum samples of different infection degree were measured, which demonstrated that the immunosensor meets the demands of clinical analysis. It exhibits some advantages, such as simplicity of fabrication, rapidity of measurement, and satisfactory sensitivity and reproducibility.

[1]  Ihab Abdel-Hamid,et al.  Development of a flow-through immunoassay system , 1998 .

[2]  Akio Tsuji,et al.  Enzymatic immunoassay of α-fetoprotein, insulin and 17-α-hydroxyprogesterone base on chemiluminescence in a flow-injection system , 1985 .

[3]  S. Alegret,et al.  Amperometric immunosensors based on rigid conducting immunocomposites. , 1997, Analytical chemistry.

[4]  Development of a sensitive, selective electrochemical immunoassay for progesterone in cow's milk based on a disposable screen-printed amperometric biosensor , 1998 .

[5]  George G. Guilbault,et al.  Amperometric immunosensor for granulocyte-macrophage colony-stimulating factor using screen-printed electrodes , 1999 .

[6]  J Wang,et al.  Electroanalysis and biosensors. , 1993, Analytical chemistry.

[7]  K R Rogers,et al.  Sol-gel-derived thick-film amperometric immunosensors. , 1998, Analytical chemistry.

[8]  K R Rogers,et al.  Thick-film electrochemical immunosensor based on stripping potentiometric detection of a metal ion label. , 1998, Analytical chemistry.

[9]  J. Park,et al.  Disposable liposome immunosensor for theophylline combining an immunochromatographic membrane and a thick-film electrode , 1999 .

[10]  N. Blanckaert,et al.  Detection and classification of paraproteins by capillary immunofixation/subtraction. , 1998, Clinical chemistry.

[11]  O. Sadik,et al.  Pulsed electrochemical technique for monitoring antibody-antigen reactions at interfaces , 1998 .

[12]  D Rodbard,et al.  Statistical quality control and routine data processing for radioimmunoassays and immunoradiometric assays. , 1974, Clinical chemistry.

[13]  G. Wallace,et al.  Determination of P-Cresol (and Other Phenolics) Using a Conducting Polymer Based Electro-Immunological Sensing System , 1994 .

[14]  C. Padeste,et al.  Amperometric immunosensing using microperoxidase MP-11 antibody conjugates , 1998 .

[15]  R. Hawkes,et al.  A dot-immunobinding assay for monoclonal and other antibodies. , 1982, Analytical biochemistry.

[16]  Damià Barceló,et al.  Reversible immunosensor for the automatic determination of atrazine. Selection and performance of three polyclonal antisera , 1999 .

[17]  E. Fábregas,et al.  Determination of β-HCG using amperometric immunosensors based on a conducting immunocomposite , 1999 .