Issue 20, 2012
From the journal:

Analyst

Ultrasensitive and selective homogeneous sandwich immunoassay detection by Surface Enhanced Raman Scattering (SERS)†

Mustafa Ersin Pekdemir,a   Deniz Ertürkan,b   Haluk Külah,c   İsmail H. Boyacı,d   Canan Özgenb  and  Uğur Tamer*a  

* Corresponding authors

a Department of Analytical Chemistry, Faculty of Pharmacy, Gazi University, Etiler, Golbasi, Ankara, Turkey, Turkey
E-mail: utamer@gazi.edu.tr

b Department of Chemical Engineering, Middle East Technical University, Ankara, Turkey

c Department of Electrical and Electronics Engineering, Middle East Technical University, Metu, Ankara, Turkey, Turkey

d Department of Food Engineering, Hacettepe University, Beytepe, Ankara, Turkey

Abstract

In this report, a simple and highly selective homogeneous sandwich immunoassay was developed for ultrasensitive detection of Staphylococcal Enterotoxin B (SEB) using Surface Enhanced Raman Scattering (SERS). The assay uses polyclonal-antibody functionalized magnetic gold nanorod particles as capture probes for SEB, which can be collected via a simple magnet. After separating SEB from the sample matrix, they are sandwiched by using binding-specific antibody–antigen pairs with the help of gold nanorod particles. Gold nanorod particles are bifunctional by design and contain self-assembled monolayers (SAMs) of a SERS tag molecule (5,5-dithiobis (2-nitrobenzoic acid), DTNB) and carboxylic functionalities of DTNB for coupling with a suitable antibody. The correlation between the SEB concentration and SERS signal was found to be linear within the range of 3 fM to 0.3 μM. The limit of detection for the assay was determined to be 768 aM (ca., 9250 SEB molecules per 20 μL sample volume). The gold heterogeneous assay system for SEB detection was also compared with the same SERS probes and gold-coated surfaces as capture substrates. The developed method was further evaluated for detecting SEB in artificially contaminated milk. Finally, the method was used for investigating the SEB specificity on bovine serum albumin (BSA) and avidin.

Graphical abstract: Ultrasensitive and selective homogeneous sandwich immunoassay detection by Surface Enhanced Raman Scattering (SERS)

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Supplementary files

Article information

DOI
https://doi.org/10.1039/C2AN35471C
Article type
Paper
Submitted
09 Apr 2012
Accepted
09 Aug 2012
First published
03 Sep 2012

Analyst, 2012,137, 4834-4840

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Ultrasensitive and selective homogeneous sandwich immunoassay detection by Surface Enhanced Raman Scattering (SERS)

M. E. Pekdemir, D. Ertürkan, H. Külah, İ. H. Boyacı, C. Özgen and U. Tamer, Analyst, 2012, 137, 4834 DOI: 10.1039/C2AN35471C

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