Applied Biochemistry and Biotechnology

, Volume 187, Issue 2, pp 493–505 | Cite as

Rapid and Sensitive Detection of Staphylococcal Enterotoxin B by Recombinant Nanobody Using Phage Display Technology

  • Saeed Zanganeh
  • Hamideh Rouhani NejadEmail author
  • Jalil Fallah Mehrabadi
  • Razieh Hosseini
  • Bahareh Shahi
  • Zahra Tavassoli
  • Asieh Aramvash


Staphylococcal enterotoxin B, from Staphylococcus aureus (S. aureus), is one of the most potent bacterial superantigens with profound toxic effects on the immune system. It is associated with food poisoning, toxic shock, atopic dermatitis, asthma, and nasal polyps in humans. The current diagnostic methods for staphylococcal enterotoxin are mainly based on traditional monoclonal antibodies which hardly meet the requirements for clinical applications, and hybridoma clones lose their ability to secrete antibodies during time. The present study investigates the development of a novel, highly specific, low-cost, and sensitive nanobody capable of being used in immunoassays for Staphylococcal enterotoxin B (SEB) detection in suspicious foods. For this purpose, Camelus dromedarius was immunized against SEB toxin. After obtaining acceptable titration, a high-quality phage display nanobody library (4 × 1010 PFU/ml) was constructed. High-affinity SEB-specific nanobodies were retrieved from constructed libraries. After phage rescue and five round of biopanning, clone screening was performed by phage ELISA. Recombinant nanobodies which were expressed from C7 and C21 clone showed the highest affinity for SEB. The presence of high quality and pure nanobody band at ~ 15 kDa was confirmed by SDS-PAGE and western blotting. The affinity constant which was measured by ELISA was calculated to be around 10−9 M. The results suggest that the proposed detection method by nanobodies is an alternative diagnostic tool enabling a rapid, inexpensive, and specific detection of the SEB.


Camel Nanobody Single-domain antibody Phage display VHH Staphylococcal enterotoxin B 



Camelid heavy chain antibody


Enzyme-linked immunoabsorbent immunoassay


Heavy chain antibodies


Staphylococcal enterotoxin B


Sodium dodecyl sulfate polyacryl amide gel electrophoresis



The authors would like to thank the research council of Malek-Ashtar University of Technology for the financial support of this investigation.

Compliance with Ethical Standards

Conflict of Interest

The authors declare no conflict of interest.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Saeed Zanganeh
    • 1
  • Hamideh Rouhani Nejad
    • 2
    Email author
  • Jalil Fallah Mehrabadi
    • 3
  • Razieh Hosseini
    • 2
  • Bahareh Shahi
    • 2
  • Zahra Tavassoli
    • 2
  • Asieh Aramvash
    • 2
  1. 1.Biotechnology Research CenterPasteur Institute of IranTehranIran
  2. 2.Malek-Ashtar University of TechnologyTehranIran
  3. 3.The Lister Laboratory of MicrobiologyTehranIran

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