Journal
ANALYTICAL CHEMISTRY
Volume 82, Issue 4, Pages 1527-1534Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac902768f
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Funding
- National Basic Research Program of China [2010CB732403]
- National Natural Science Foundation of China [20877019, 20735002]
- Key Natural Sciences Foundation of Fujian Province, China [D0520001]
- Key Program of Science and Technology Department of Fujian Province, China [2007Y0026]
- NTU-MOE Academic Research Funds [RG65/08]
- High-Qualified Talent Funding of Fuzhou University [0460-022275]
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A novel nanoparticle-based electrochemical immunoassay of carbohydrate antigen 125 (CA125) as a model was designed to couple with a microfluidic strategy using anti-CA125-functionalized magnetic beads as immunosensing probes. To construet the immunoassay,thionine-horseradish peroxidase conjugation (TH-HRP) was initially doped into nanosilica particles using the reverse micelle method, and then HRP-labeled anti-CA125 antibodies (HRP-anti-CA125) were bound onto the surface of the synthesized nanoparticles, which were used as recognition elements. Different from conventional nanoparticle-based electrochemical immunoassays, the recognition elements of the immunoassay simultaneously contained electron mediator and enzyme labels and simplified the electrochemical measurement process. The sandwich-type immunoassay format was used for the online formation of the immuno-complex in an incubation cell and captured in the detection cell with an external magnet. The electrochemical signals derived from the carried HRP toward the reduction of H2O2 using the doped thionine as electron mediator. Under optimal conditions, the electrochemical immunoassay exhibited a wide working range from 0.1 to 450 U/mL with a detection limit of 0.1 U/mL CA125. The precision, reproducibility, and stability of the immunoassay were acceptable. The assay was evaluated for clinical serum samples, receiving in excellent accordance with results obtained from the standard enzyme-linked immunosorbent assay (ELISA) method. Concluding, the nanoparticle-based assay format provides a promising approach in clinical application and thus represents a versatile detection method.
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