Issue 1, 2004
From the journal:

Analyst

Immunoassay using probe-labelling immunogold nanoparticles with silver staining enhancement via surface-enhanced Raman scattering

Shuping Xu,a   Xiaohui Ji,b   Weiqing Xu,*a   Xiaoling Li,a   Lianying Wang,b   Yubai Bai,*b   Bing Zhaoa  and  Yukihiro Ozakic  

* Corresponding authors

a Key Laboratory for Supramolecular Structure and Material of Ministry of Education, Jilin University, Changchun 130021, P. R. China

b College of Chemistry, Jilin University, Changchun 130021, P. R. China
E-mail: wqxu@jlu.edu.cn
Fax: +86-431-8980729
Tel: +86-431-8498912

c Department of Chemistry, School of Science and Technology, Kwansei-Gakuin University, Sanda, Hyogo 669-1337, Japan

Abstract

This paper reports a novel immunoassay based on surface-enhanced Raman scattering (SERS) and immunogold labelling with silver staining enhancement. Immunoreactions between immunogold colloids modified by a Raman-active probe molecule (e.g., 4-mercaptobenzoic acid) and antigens, which were captured by antibody-assembled chips such as silicon or quartz, were detected via SERS signals of Raman-active probe molecule. All the self-assembled steps were subjected to the measurements of ultraviolet-visible (UV-vis) spectra to monitor the formation of a sandwich structure onto a substrate. The immunoassay was performed by a sandwich structure consisting of three layers. The first layer was composed of immobilized antibody molecules of mouse polyclonal antibody against Hepatitis B virus surface antigen (PAb) on a silicon or quartz substrate. The second layer was the complementary Hepatitis B virus surface antigen (Antigen) molecules captured by PAb on the substrate. The third layer was composed of the probe-labelling immunogold nanoparticles, which were modified by mouse monoclonal antibody against Hepatitis B virus surface antigen (MAb) and 4-mercaptobenzoic acid (MBA) as the Raman-active probe on the surface of gold colloids. After silver staining enhancement, the antigen is identified by a SERS spectrum of MBA. A working curve of the intensity of a SERS signal at 1585 cm−1 due to the ν8a aromatic ring vibration of MBA versus the concentration of analyte (Antigen) was obtained and the non-optimized detection limit for the Hepatitis B virus surface antigen was found to be as low as 0.5 µg mL−1.

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Article information

DOI
https://doi.org/10.1039/B313094K
Article type
Paper
Submitted
17 Oct 2003
Accepted
01 Dec 2003
First published
11 Dec 2003

Analyst, 2004,129, 63-68

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Immunoassay using probe-labelling immunogold nanoparticles with silver staining enhancement via surface-enhanced Raman scattering

S. Xu, X. Ji, W. Xu, X. Li, L. Wang, Y. Bai, B. Zhao and Y. Ozaki, Analyst, 2004, 129, 63 DOI: 10.1039/B313094K

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