Nanobody-based enzyme immunoassay for ochratoxin A in cereal with high resistance to matrix interference

Xing Liu, Zongwen Tang, Zhenhua Duan, Zhenyun He, Mei Shu, Xianxian Wang, Shirley J. Gee, Bruce D. Hammock, Yang Xu

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

A sensitive indirect competitive nanobody-based enzyme linked immunosorbent assay (Nb-ELISA) for ochratoxin A (OTA) with high resistance to cereal matrix interference was developed. Nanobodies against OTA (Nb15, Nb28, Nb32, Nb36) were expressed in E. coli cells and their thermal stabilities were compared with that of an OTA-specific monoclonal antibody 6H8. All nanobodies could still retain their antigen-binding activity after exposure to temperature 95 °C for 5 min or to 90 °C for 75 min. Nb28 that exhibited the highest sensitivity in ELISA was selected for further research. An indirect competitive ELISA based on Nb28 was developed for OTA, with an IC50 of 0.64 ng/mL and a linear range (IC20-IC80) of 0.27–1.47 ng/mL. Cereal samples were analyzed following a 2.5 fold dilution of sample extracts, showing the good resistance to matrix interference of the Nb-ELSIA. The recovery of spiked cereal samples (rice, oats, barley) ranged from 80% to 105% and the Nb-ELISA results of OTA content in naturally contamined samples were in good agreement with those determined by a commercial ELISA kit. The results indicated the reliablity of nanobody as a promising immunoassay reagent for detection of mycotoxins in food matrix and its potential in biosensor development.

Original languageEnglish (US)
Pages (from-to)154-158
Number of pages5
JournalTalanta
Volume164
DOIs
StatePublished - Mar 1 2017

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Single-Domain Antibodies
Immunoenzyme Techniques
Enzyme-Linked Immunosorbent Assay
Enzymes
Immunosorbents
Assays
Mycotoxins
Biosensing Techniques
Hordeum
Immunoassay
Biosensors
Escherichia coli
Dilution
Inhibitory Concentration 50
Thermodynamic stability
Hot Temperature
Monoclonal Antibodies
Edible Grain
ochratoxin A
Antigens

Keywords

  • Cereal
  • Immunoassay
  • Matrix interference
  • Nanobody
  • Ochratoxin A

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Nanobody-based enzyme immunoassay for ochratoxin A in cereal with high resistance to matrix interference. / Liu, Xing; Tang, Zongwen; Duan, Zhenhua; He, Zhenyun; Shu, Mei; Wang, Xianxian; Gee, Shirley J.; Hammock, Bruce D.; Xu, Yang.

In: Talanta, Vol. 164, 01.03.2017, p. 154-158.

Research output: Contribution to journalArticle

Liu, X, Tang, Z, Duan, Z, He, Z, Shu, M, Wang, X, Gee, SJ, Hammock, BD & Xu, Y 2017, 'Nanobody-based enzyme immunoassay for ochratoxin A in cereal with high resistance to matrix interference', Talanta, vol. 164, pp. 154-158. https://doi.org/10.1016/j.talanta.2016.11.039
Liu, Xing ; Tang, Zongwen ; Duan, Zhenhua ; He, Zhenyun ; Shu, Mei ; Wang, Xianxian ; Gee, Shirley J. ; Hammock, Bruce D. ; Xu, Yang. / Nanobody-based enzyme immunoassay for ochratoxin A in cereal with high resistance to matrix interference. In: Talanta. 2017 ; Vol. 164. pp. 154-158.
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