Journal
TALANTA
Volume 123, Issue -, Pages 161-168Publisher
ELSEVIER
DOI: 10.1016/j.talanta.2014.02.015
Keywords
Immunoassay; Surface-enhanced Raman scattering; Biosensing; Nanoparticle; SERS substrate
Categories
Funding
- National Natural Science Foundation of China [61036013, 61138003, 61275153, 61265009, 61320106014]
- Natural Science Foundation of Zhejiang [LY12A04002]
- International Collaboration Program of the Natural Science Foundation of Ningbo [2010D10018, 2012A610107]
- K.C. Wong Education Foundation
- K.C. Wong Magna Foundation of Ningbo University, China
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A super-high-sensitivity immunoassay based on surface-enhanced Raman scattering (SERS) was implemented using the nano-Au immune probes and nano-Ag immune substrate. Ultraviolet-visible extinction spectra, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) images, and SERS spectra were used to characterise the nano-Au immune probes and the nano-Ag immune substrate. The nano-Ag immune substrate was prepared by the in situ growth of Ag nanoparticles and the subsequent linkage of these nanoparticles with anti-apolipoprotein B on a silicon wafer. The nano-Ag immune substrate exhibited strong SERS activity, excellent reproducibility, and high biospecificity. The nano-Au immune probes were prepared by immobilising 4-mercaptobenzoic acid (4MBA) molecules as a Raman reporter and anti-apolipoprotein B onto the surfaces of Au nanoparticles. It was found that 4MBA induced the aggregation of Au nanoparticles, resulting in the generation of vast hot spots. Moreover, the nano-Au immune probes exhibited strong SERS activity and high biospecificity. A sandwich-type immunoassay structure consisting of the nano-Au immune probes and nano-Ag immune substrate was used to detect the concentration of apolipoprotein B, where the detection limit was as low as 2 fg/mL (3.878 x 10(-18) mol/L). Taken together, the experimental results indicate that the proposed immunoassay protocol has a great potential application in biological sensing and clinical diagnostics. (c) 2014 Elsevier B.V. All rights reserved.
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