Competitive and noncompetitive immunoassays for the detection of benzothiostrobin using magnetic nanoparticles and fluorescein isothiocyanate-labeled peptides

  • He Chen
  • Qian Yang
  • Yuan Ding
  • Natalia Vasylieva
  • Candace S. Bever
  • Xiude Hua
  • Minghua Wang
  • Bruce D. Hammock
Research Paper


Phage-displayed peptides have been proven to be powerful reagents for competitive and noncompetitive immunoassays. However, they are unconventional reagents, which greatly limit their analytical commercial applications and require additional reagents for detection. In this work, the peptides that specifically bind with anti-benzothiostrobin monoclonal antibody (mAb) or benzothiostrobin-mAb immunocomplex were synthesized and conjugated with fluorescein isothiocyanate (FITC) as substitutes of the phage-displayed peptides to avoid their shortcomings and extend their applications. Competitive and noncompetitive fluorescence immunoassays (FIAs) for benzothiostrobin were developed by mAb coupling with magnetic nanoparticles as concentration elements and peptides conjugated with FITC as tracers. Compared with enzyme-linked immunosorbent assays, the FIAs reduced the number of steps from 6 to 2 and analysis time from more than 5 to 1.2 h. The competitive FIA showed the half-maximal inhibition concentration (IC50) of 16.8 ng mL−1 and detection range (IC10–IC90) of 1.0–759.9 ng mL−1, while the concentration of analyte producing 50% saturation of the signal (SC50) and detection range (SC10–SC90) of noncompetitive FIA were 93.4 and 5.9–788.2 ng mL−1, respectively. The average spiked recoveries were 68.33–98.50% and 73.33–96.67% for competitive and noncompetitive FIAs, respectively. The FIAs showed good correlation with high-performance liquid chromatography for the detection of benzothiostrobin in authentic samples.

Graphical abstract

Development of competitive and noncompetitive fluorescence immunoassays for benzothiostrobin by using monoclonal antibody coupling with magnetic nanoparticles as concentration elements and peptides conjugated with fluorescein isothiocyanate as tracers.


Pesticide residue Immunoassay Peptidomimetic Immunocomplex Benzothiostrobin 



This work was funded by the National Key Research and Development Program of China (2017YFF0210200), the Fundamental Research Funds for the Central Universities (KYZ201618), and the National Institute of Environmental Health Sciences, Superfund Research Program (P42 ES04699). C.S.B. was funded by the United States Department of Agriculture, Agricultural Research Service, National Program project NP108, CRIS 5325-42000-049-00D, and by an interagency agreement IAA # 60-2030-5-004 with the Department of Homeland Security.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2018_1478_MOESM1_ESM.pdf (815 kb)
ESM 1 (PDF 814 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • He Chen
    • 1
  • Qian Yang
    • 1
  • Yuan Ding
    • 1
  • Natalia Vasylieva
    • 2
  • Candace S. Bever
    • 3
  • Xiude Hua
    • 1
    • 2
  • Minghua Wang
    • 1
  • Bruce D. Hammock
    • 2
  1. 1.College of Plant Protection (State & Local Joint Engineering Research Center of Green Pesticide Invention and Application)Nanjing Agricultural UniversityNanjingChina
  2. 2.Department of Entomology and UCD Cancer CenterUniversity of CaliforniaDavisUSA
  3. 3.Foodborne Toxin Detection and Prevention Unit, Agricultural Research Service, United States Department of AgricultureWestern Regional Research CenterAlbanyUSA

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