TY - JOUR
T1 - Enzyme-catalyzed silver deposition on irregular-shaped gold nanoparticles for electrochemical immunoassay of alpha-fetoprotein
AU - Lai, Wenqiang
AU - Tang, Dianping
AU - Que, Xiaohua
AU - Zhuang, Junyang
AU - Fu, Libing
AU - Chen, Guonan
PY - 2012/11/28
Y1 - 2012/11/28
N2 - A new and disposable electrochemical immunosensor was designed for detection of alpha-fetoprotein (AFP), as a model analyte, with sensitivity enhancement based on enzyme-catalyzed silver deposition onto irregular-shaped gold nanoparticles (ISGNPs). The assay was carried out with a sandwich-type immunoassay protocol by using ISGNP-labeled anti-AFP antibodies conjugated with alkaline phosphatase (ALP-Ab2) as detection antibodies. The enzymatically catalytic deposition of silver on the electrode could be measured by stripping analysis in KCl solution due to the Ag/AgCl solid-state voltammetric process. Several labeling protocols including spherical gold nanoparticle-labeled ALP-Ab2 and ISGNP-labeled ALP-Ab2 were investigated for determination of AFP, and improved analytical properties were achieved with the ISGNP labeling. With the ISGNP labeling method, the effects of incubation time and incubation temperature for antigen-antibody reaction, and deposition time of silver on the current responses of the electrochemical immunosensors were also monitored. Under optimal conditions, the electrochemical immunosensor exhibited a wide dynamic range from 0.01ngmL-1 to 200ngmL-1 with a detection limit of 5.0pgmL-1 AFP. The immunosensor displayed a good stability and acceptable reproducibility and accuracy. No significant differences at the 95% confidence level were encountered in the analysis of 10 clinical serum samples between the developed immunoassay and the commercially available electrochemiluminescent method for determination of AFP.
AB - A new and disposable electrochemical immunosensor was designed for detection of alpha-fetoprotein (AFP), as a model analyte, with sensitivity enhancement based on enzyme-catalyzed silver deposition onto irregular-shaped gold nanoparticles (ISGNPs). The assay was carried out with a sandwich-type immunoassay protocol by using ISGNP-labeled anti-AFP antibodies conjugated with alkaline phosphatase (ALP-Ab2) as detection antibodies. The enzymatically catalytic deposition of silver on the electrode could be measured by stripping analysis in KCl solution due to the Ag/AgCl solid-state voltammetric process. Several labeling protocols including spherical gold nanoparticle-labeled ALP-Ab2 and ISGNP-labeled ALP-Ab2 were investigated for determination of AFP, and improved analytical properties were achieved with the ISGNP labeling. With the ISGNP labeling method, the effects of incubation time and incubation temperature for antigen-antibody reaction, and deposition time of silver on the current responses of the electrochemical immunosensors were also monitored. Under optimal conditions, the electrochemical immunosensor exhibited a wide dynamic range from 0.01ngmL-1 to 200ngmL-1 with a detection limit of 5.0pgmL-1 AFP. The immunosensor displayed a good stability and acceptable reproducibility and accuracy. No significant differences at the 95% confidence level were encountered in the analysis of 10 clinical serum samples between the developed immunoassay and the commercially available electrochemiluminescent method for determination of AFP.
KW - Alpha-fetoprotein
KW - Electrochemical immunosensor
KW - Enzyme-catalyzed silver deposition
KW - Irregular-shaped gold nanoparticles
UR - http://www.scopus.com/inward/record.url?scp=84869085744&partnerID=8YFLogxK
U2 - 10.1016/j.aca.2012.10.028
DO - 10.1016/j.aca.2012.10.028
M3 - Article
C2 - 23146395
AN - SCOPUS:84869085744
SN - 0003-2670
VL - 755
SP - 62
EP - 68
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
ER -