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Naked-eye sensitive ELISA-like assay based on gold-enhanced peroxidase-like immunogold activity

Title
Naked-eye sensitive ELISA-like assay based on gold-enhanced peroxidase-like immunogold activity
Author
주재범
Keywords
Gold nanoparticles; Peroxidase-like activity; Colorimetric immunoassay; Human IgG; Gold enhancement
Issue Date
2016-01
Publisher
SPRINGER HEIDELBERG
Citation
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, v. 408, NO 4, Page. 1015-1022
Abstract
A naked-eye sensitive ELISA-like assay was developed based on gold-enhanced peroxidase-like activity of gold nanoparticles (AuNPs). Using human IgG (H-IgG) as an analytical model, goat anti-human IgG antibody (anti-IgG) adsorbed on microtiter plate and AuNPs-labeled anti-IgG acted as capture antibody and detection antibody, respectively. Because the surfaces of AuNPs were blocked by protein molecules, the peroxidase-like activity of AuNPs was almost inhibited, evaluated by the catalytic oxidation of peroxidase enzyme substrate 3,3',5,5'-tetramethylbenzidine (TMB), which could produce a bright blue color in the presence of H2O2. Fortunately, the catalytic ability of AuNPs was dramatically increased by the deposition of gold due to the formation of a new gold shell on immunogold. Under optimal reaction conditions, the colorimetric immunoassay presented a good linear relationship in the range of 0.7-100 ng/mL and the limit of detection (LOD) of 0.3 ng/mL calculated by 3 sigma/S for UV-vis detection, and obtained LOD of 5 ng/mL for naked-eye detection. The obtained results were competitive with conventional sandwich ELISA with the LOD of 1.6 ng/mL. Furthermore, this developed colorimetric immunoassay was successfully applied to diluted human serum and fetal bovine serum samples, and predicted a broad prospect for the use of peroxidase-like activity involving nanomaterials in bioassay and diagnostics.
URI
https://link.springer.com/article/10.1007%2Fs00216-015-9219-8http://hdl.handle.net/20.500.11754/31446
ISSN
1618-2642; 1618-2650
DOI
10.1007/s00216-015-9219-8
Appears in Collections:
GRADUATE SCHOOL[S](대학원) > BIONANOTECHNOLOGY(바이오나노학과) > Articles
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