Analytical and Bioanalytical Chemistry

, Volume 409, Issue 26, pp 6227–6234 | Cite as

Gold nanoparticle-based localized surface plasmon immunosensor for staphylococcal enterotoxin A (SEA) detection

  • Maroua Ben Haddada
  • David Hu
  • Michèle Salmain
  • Lu Zhang
  • Chen Peng
  • Yi Wang
  • Bo Liedberg
  • Souhir BoujdayEmail author
Research Paper


We describe the engineering of stable gold nanoparticle (AuNP) bioconjugates for the detection of staphylococcal enterotoxin A (SEA) using localized surface plasmon resonance (LSPR). Two types of AuNP bioconjugates were prepared by covalently attaching anti-SEA antibody (Ab) or SEA to AuNPs. This was achieved by reacting Traut’s reagent with lysine residues of both proteins to generate thiol groups that bind to gold atoms on the AuNP surface. These bioconjugates were characterized in-depth by absorption spectroscopy, cryo-transmission electron microscopy, dynamic light scattering, and zeta potential measurements. Their stability over time was assessed after 1 year storage in the refrigerator at 4 °C. Two formats of homogeneous binding assays were set up on the basis of monitoring of LSPR peak shifts resulting from the immunological reaction between the (i) immobilized antibody and free SEA, the direct assay, or (ii) immobilized SEA and free antibody, the competitive assay. In both formats, a correlation between the LSPR band shift and SEA concentration could be established. Though the competitive format did not meet the expected analytical performance, the direct format, the implementation of which was very simple, afforded a specific and sensitive response within a broad dynamic range—nanogram per milliliter to microgram per milliliter. The limit of detection (LOD) of SEA was estimated to equal 5 ng/mL, which was substantially lower than the LOD obtained using a quartz crystal microbalance. Moreover, the analytical performance of AuNP-Ab bioconjugate was preserved after 1 year of storage at 4 °C. Finally, the LSPR biosensor was successfully applied to the detection of SEA in milk samples. The homogeneous nanoplasmonic immunosensor described herein provides an attractive alternative for stable and reliable detection of SEA in the nanogram per milliliter range and offers a promising avenue for rapid, easy to implement, and sensitive biotoxin detection.

Sensitive LSPR Biosensing of SEA in buffer and milk using stable AuNP-Antibody bioconjugates

Graphical abstract


Immunosensor Localized surface plasmon resonance Staphylococcal enterotoxin A Gold nanoparticles 



We would like to thank the DIM Analytics and Region Ile-de-France for M. Ben Haddada PhD scholarship. We also thank Anton Paar for the access to Litesizer™ 500 apparatus. This work was supported by the iFood initiative Nanyang Technological University, by the French-Singaporean PHC Merlion program (grant 5.03.15), and by the ANR-FWF program (grant ANR-15-CE29-0026).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

216_2017_563_MOESM1_ESM.pdf (633 kb)
ESM 1 (PDF 633 kb)


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Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • Maroua Ben Haddada
    • 1
    • 2
  • David Hu
    • 1
  • Michèle Salmain
    • 2
  • Lu Zhang
    • 1
    • 3
    • 4
  • Chen Peng
    • 3
  • Yi Wang
    • 3
  • Bo Liedberg
    • 3
  • Souhir Boujday
    • 1
    • 4
    Email author
  1. 1.UPMC Univ Paris 6, CNRS, Laboratoire de Réactivité de Surface (LRS)Sorbonne UniversitésParisFrance
  2. 2.UPMC Univ Paris 06, CNRS, Institut Parisien de Chimie Moléculaire (IPCM)Sorbonne UniversitésParisFrance
  3. 3.Centre for Biomimetic Sensor Science, School of Material Science and EngineeringNanyang Technological UniversityNanyangSingapore
  4. 4.MajuLab, UMI 3654, CNRS-UNS-NUS-NTU International Joint Research UnitNanyangSingapore

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