期刊
MICROCHIMICA ACTA
卷 179, 期 1-2, 页码 71-81出版社
SPRINGER WIEN
DOI: 10.1007/s00604-012-0870-x
关键词
Nanoporous gold; Immunoassay; Self-assembled monolayer; Square-wave voltammetry; Carcinoembryonic antigen; Prostate specific antigen
资金
- UM-St. Louis
- NIGMS [R01-GM090254]
Nanoporous gold (NPG) was utilized as a support for immobilizing alkaline phosphatase (ALP) conjugated to monoclonal antibodies against either prostate specific antigen (PSA) or carcinoembryonic antigen (CEA). The antibody-ALP conjugates were coupled to self-assembled monolayers of lipoic acid and used in direct kinetic assays. Using the enzyme substrate p-aminophenylphosphate, the product p-aminophenol was detected by its oxidation near 0.1 V (vs. Ag|AgCl) using square wave voltammetry. The difference in peak current arising from oxidation of p-aminophenol before and after incubation with biomarker increased with biomarker concentration. The response to these two biomarkers was linear up to 10 ng mL(-1) for CEA and up to 30 ng mL(-1) for PSA. The effect of interference on the PSA assay was studied using bovine serum albumin (BSA) as a model albumin protein. The effect of interference from a serum matrix was examined for the PSA assay using newborn calf serum. A competitive version of the immunoassay using antigen immobilized onto the NPG surface was highly sensitive at lower antigen concentration. Estimates of the surface coverage of the antibody-ALP conjugates on the NPG surface are presented.
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