A disposable multianalyte electrochemical immunosensor array for automated simultaneous determination of tumor markers.
BACKGROUND
Automated and convenient multianalyte detection with high throughput is increasingly needed in clinical diagnosis. We developed a disposable 4-by-2 array for programmed simultaneous amperometric immunoassay of 4 tumor markers.


METHODS
We used a screen-printed technique, 1-step immobilization method, and flow injection technique. We immobilized carcinoembryonic antigen, alpha-fetoprotein, beta-human choriogonadotropin, and carcinoma antigen 125 as model analytes in a redox mediator-grafted, biopolymer-modified, screen-printed carbon electrode array to capture corresponding horseradish peroxidase-labeled antibodies in competitive immunoreactions. The simultaneous multianalyte immunoassay was automatically carried out to amperometrically monitor the mediator-catalyzed enzymatic response to hydrogen peroxide, which decreased in proportion to the concentrations of analytes in samples.


RESULTS
The multianalyte immunosensor array had a throughput of 60 samples/h and allowed simultaneous detection of carcinoembryonic antigen, alpha-fetoprotein, beta-human choriogonadotropin, and carcinoma antigen 125 in clinical serum samples with concentrations up to 188 microg/L, 250 microg/L, 266 IU/L, and 334 kIU/L, respectively. The detection limits (limits of the blank, mean of blank plus 3 SD) were 1.1 micro/L, 1.7 microg/L, 1.2 IU/L, and 1.7 kIU/L. The inter- and intraassay imprecision (CVs) of the immunosensor arrays were <7.8% and <9.0%, respectively. The immunosensor arrays were stable for 28 days.


CONCLUSIONS
This newly constructed immunosensor array provides a simple, automated, simultaneous multianalyte immunoassay with high throughput, short analytical time, and sufficiently low detection limits for clinical application. This method offers the capability of miniaturizing the multianalyte detection device.
Wu
Jie
J
Key Laboratory of Analytical Chemistry for Life Science (Ministry of Education of China), Department of Chemistry, Nanjing University, Nanjing 210093, Peoples Republic of China.
Yan
Feng
F
Tang
Jinhai
J
Zhai
Chun
C
Ju
Huangxian
H
D001322:Autoanalysis; D014408:Biomarkers, Tumor / Q000097:blood*; D018394:CA-125 Antigen / Q000097:blood; D002272:Carcinoembryonic Antigen / Q000097:blood; D048271:Chitosan / Q000737:chemistry; D006063:Chorionic Gonadotropin / Q000097:blood; D003429:Cross Reactions; D004563:Electrochemistry; D006801:Humans; D007118:Immunoassay; D007202:Indicators and Reagents; D046228:Microarray Analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014048:Tolonium Chloride / Q000737:chemistry; D000509:alpha-Fetoproteins / Q000032:analysis
D014408:Biomarkers, Tumor; D018394:CA-125 Antigen; D002272:Carcinoembryonic Antigen; D006063:Chorionic Gonadotropin; D007202:Indicators and Reagents; D000509:alpha-Fetoproteins; D014048:Tolonium Chloride; D048271:Chitosan