Advertisement

Analytical and Bioanalytical Chemistry

, Volume 411, Issue 6, pp 1287–1295 | Cite as

Development of a one-step immunoassay for triazophos using camel single-domain antibody–alkaline phosphatase fusion protein

  • Kai Wang
  • Zhiping Liu
  • Guochun DingEmail author
  • Ji Li
  • Natalia Vasylieva
  • Qing X. Li
  • Dongyang Li
  • Shirley J. Gee
  • Bruce D. Hammock
  • Ting XuEmail author
Research Paper
  • 28 Downloads

Abstract

Triazophos is mainly used in Asian and African countries for the control of insects in agricultural production. Camelid variable domains of heavy-chain antibodies (VHHs) show great promise in monitoring environmental chemicals such as pesticides. To improve the rate of success in the generation of VHHs against triazophos, genes specifically encoding VHH fragments from the unique allotype IgG3a of an immunized Camelus bactrianus were amplified by using a pair of novel primers and introduced to construct a diverse VHH library. Five out of seven isolated positive clones, including the VHH T1 with the highest affinity to triazophos, were derived from the allotype IgG3a. A one-step enzyme-linked immunosorbent assay (ELISA) using VHH T1 genetically fused with alkaline phosphatase (AP) had a half-maximum inhibition concentration of 6.6 ng/mL for triazophos. This assay showed negligible cross-reactivity with a list of important organophosphate pesticides (< 0.1%). The average recoveries of triazophos from water, soil, and apple samples determined by the one-step ELISA ranged from 83 to 108%, having a good correlation with those by a gas chromatography mass spectrometry (R2 = 0.99). The VHH-AP fusion protein shows potential for the analysis of triazophos in various matrices.

Keywords

Variable domains of heavy-chain antibody IgG3a VHH-AP One-step ELISA Triazophos 

Notes

Acknowledgements

The content is solely the responsibility of the authors and does not necessarily represent the official views of the funding agencies.

Funding information

This work was supported in part by the Key Project of Inter-Governmental International Scientific and Technological Innovation Cooperation (2016YFE0108900), the National Key Research and Development Program of China (2016YFD0800606), and the National Institute of Environmental Health Sciences Superfund Research Program (P42ES04699), USA.

Compliance with ethical standards

The animal experiments were approved by the China Agricultural University Animal Care and Use Committee. All procedures performed in this research involving Bactrian camels were in accordance with the ethical standards of the China Agricultural University Animal Care and Use Committee. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2018_1563_MOESM1_ESM.pdf (574 kb)
ESM 1 (PDF 574 kb)

References

  1. 1.
    Hamers-Casterman C, Atarhouch T, Muyldermans S, Robinson G, Hammers C, Songa EB, et al. Naturally occurring antibodies devoid of light chains. Nature. 1993;363(6428):446–8.CrossRefGoogle Scholar
  2. 2.
    Bever CS, Dong JX, Vasylieva N, Barnych B, Cui Y, Xu ZL, et al. VHH antibodies: emerging reagents for the analysis of environmental chemicals. Anal Bioanal Chem. 2016;408(22):5985–6002.CrossRefGoogle Scholar
  3. 3.
    Gonzalez-Sapienza G, Rossotti MA, Tabares-da Rosa S. Single-domain antibodies as versatile affinity reagents for analytical and diagnostic applications. Front Immunol. 2017;8:977.CrossRefGoogle Scholar
  4. 4.
    Wang J, Majkova Z, Bever CR, Yang J, Gee SJ, Li J, et al. One-step immunoassay for tetrabromobisphenol A using a camelid single domain antibody-alkaline phosphatase fusion protein. Anal Chem. 2015;87:4741–8.CrossRefGoogle Scholar
  5. 5.
    Shu M, Xu Y, Liu X, Li Y, He Q, Tu Z, et al. Anti-idiotypic nanobody-alkaline phosphatase fusion proteins: development of a one-step competitive enzyme immunoassay for fumonisin B1 detection in cereal. Anal Chim Acta. 2016;924:53–9.CrossRefGoogle Scholar
  6. 6.
    Anderson GP, Goldman ER. TNT detection using llama antibodies and a two-step competitive fluid array immunoassay. J Immunol Methods. 2008;339(1):47–54.CrossRefGoogle Scholar
  7. 7.
    Lange IG, Daxenberger A, Meyer HH. Studies on the antibody response of Lamaglama—evaluation of the binding capacity of different IgG subtypes in ELISAs for clenbuterol and BSA. Vet Immunol Immunopathol. 2001;83(1–2):1–9.CrossRefGoogle Scholar
  8. 8.
    Wang J, Bever CR, Majkova Z, Dechant JE, Yang J, Gee SJ, et al. Heterologous antigen selection of camelid heavy chain single domain antibodies against tetrabromobisphenol A. Anal Chem. 2014;86:8296–302.CrossRefGoogle Scholar
  9. 9.
    Pírez-Schirmer M, Rossotti MA, Badagian N, Leizagoyen C, Brena BM, Gonzalez-Sapienza GG. Comparison of three anti-hapten VHH selection strategies for the development of highly sensitive immunoassays for microcystins. Anal Chem. 2017;89:6800–6.CrossRefGoogle Scholar
  10. 10.
    Blanc MR, Anouassi A, Abed MA, Tsikis G, Canepa S, Labas V, et al. A one-step exclusion-binding procedure for the purification of functional heavy-chain and mammalian-typeγ-globulins from camelid sera. Biotechnol Appl Biochem. 2009;54(4):207–12.CrossRefGoogle Scholar
  11. 11.
    Maass DR, Sepulveda J, Pernthaner A, Shoemaker CB. Alpaca (Lama pacos) as a convenient source of recombinant camelid heavy chain antibodies (VHHs). J Immunol Methods. 2007;324(1–2):13–25.CrossRefGoogle Scholar
  12. 12.
    Kim HJ, McCoy MR, Majkova Z, Dechant JE, Gee SJ, Tabares-da Rosa S, et al. Isolation of alpaca anti-hapten heavy chain single domain antibodies for development of sensitive immunoassay. Anal Chem. 2011;84:1165–71.CrossRefGoogle Scholar
  13. 13.
    Dai DJ, Lin RH, Shen Y, Shen Y, Wang HD, Zong FL. Suggestion and countermeasures of strengtherning risk management on triazophos. Pestic Sci Admin. 2017;38(9):1–8.Google Scholar
  14. 14.
    Lal R, Jat BL. Bio-efficacy of insecticides and biorationals against the incidence of whitefly, Bemisia tabaci (Genn.) and yellow mosaic virus in mungbean. Afr J Agr Res. 2015;10(10):1050–6.CrossRefGoogle Scholar
  15. 15.
    Tafuri J, Roberts J. Organophosphate poisoning. Ann Emerg Med. 1987;16(2):193–202.CrossRefGoogle Scholar
  16. 16.
    Gui WJ, Jin RY, Chen ZL, Cheng JL, Zhu GN. Hapten synthesis for enzyme-linked immunoassay of the insecticide triazophos. Anal Biochem. 2006;3571:9–14.CrossRefGoogle Scholar
  17. 17.
    Tabares-da Rosa S, Rossotti M, Carleiza C, Carrión F, Pritsch O, Ahn KC, et al. Competitive selection from single domain antibody libraries allows isolation of high-affinity antihapten antibodies that are not favored in the llama immune response. Anal Chem. 2011;83:7213–20.CrossRefGoogle Scholar
  18. 18.
    Liang CZ, Jin RY, Gui WJ, Zhu GN. Enzyme-linked immunosorbent assay based on a monoclonal antibody for the detection of the insecticide triazophos: assay optimization and application to environmental samples. Environ Sci Technol. 2007;41(19):6783–8.CrossRefGoogle Scholar
  19. 19.
    Vasylieva N, Ahn KC, Barnych B, Gee SJ, Hammock BD. Development of an immunoassay for the detection of the phenylpyrazole insecticide fipronil. Environ Sci Technol. 2015;49(16):10038–47.CrossRefGoogle Scholar
  20. 20.
    Kim EE, Wyckoff HW. Structure of alkaline phosphatases. Clin Chim Acta. 1990;186(2):175–87.CrossRefGoogle Scholar
  21. 21.
    Zhang J, Tanha J, Hirama T, Khieu NH, To R, Tong-Sevinc H, et al. Pentamerization of single-domain antibodies from phage libraries: a novel strategy for the rapid generation of high-avidity antibody reagents. J Mol Biol. 2004;335(1):49–56.CrossRefGoogle Scholar
  22. 22.
    Liu JL, Zabetakis D, Lee AB, Goldman ER, Anderson GP. Single domain antibody–alkaline phosphatase fusion proteins for antigen detection—analysis of affinity and thermal stability of single domain antibody. J Immunol Methods. 2013;393(1–2):1–7.CrossRefGoogle Scholar
  23. 23.
    Liu SH, Wang L, Wei LH. Studies on the immunoassay for triazophos. Chin J Anal Chem. 2005;33(12):1697–700.Google Scholar
  24. 24.
    Jin MJ, Shao H, Jin F, Gui WJ, Shi XM, Wang J, et al. Enhanced competitive chemiluminescent enzyme immunoassay for the trace detection of insecticide triazophos. J Food Sci. 2012;77(5):T99–T104.CrossRefGoogle Scholar
  25. 25.
    Du PF, Jin MJ, Chen G, Zhang C, Cui XY, Zhang YD, et al. Competitive colorimetric triazophos immunoassay employing magnetic microspheres and multi-labeled gold nanoparticles along with enzymatic signal enhancement. Microchim Acta. 2017;184:3705–12.CrossRefGoogle Scholar
  26. 26.
    Ladenson RC, Crimmins DL, Landt Y, Ladenson JH. Isolation and characterization of a thermally stable recombinant anti-caffeine heavy-chain antibody fragment. Anal Chem. 2006;78:4501–8.CrossRefGoogle Scholar
  27. 27.
    Alvarez-Rueda N, Behar G, Ferré V, Pugniere M, Roquet F, Gastinel L, et al. Generation of llama single-domain antibodies against methotrexate, a prototypical hapten. Mol Immunol. 2007;44(7):1680–90.CrossRefGoogle Scholar
  28. 28.
    Bever CS, Majkova Z, Radhakrishnan R, Suni I, McCoy M, Wang Y, et al. Development and utilization of camelid VHH antibodies from alpaca for 2,2′,4,4′-tetrabrominated diphenyl ether detection. Anal Chem. 2014;86:7875–82.CrossRefGoogle Scholar
  29. 29.
    GB 2763-2016, national food safety standard maximum residue limits for pesticides in food. National standards of the People’s Republic of China.Google Scholar
  30. 30.
    Chen G, Yang LH, Jin MJ, Du PF, Zhang C, Wang J, et al. The rapid screening of triazophos residues in agricultural products by chemiluminescent enzyme immunoassay. PLoS One. 2015;10(7):e0133839.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Kai Wang
    • 1
  • Zhiping Liu
    • 1
  • Guochun Ding
    • 1
    Email author
  • Ji Li
    • 1
  • Natalia Vasylieva
    • 2
  • Qing X. Li
    • 3
  • Dongyang Li
    • 2
  • Shirley J. Gee
    • 2
  • Bruce D. Hammock
    • 2
  • Ting Xu
    • 1
    Email author
  1. 1.Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Resources and Environmental SciencesChina Agricultural UniversityBeijingChina
  2. 2.Department of Entomology and UCD Comprehensive Cancer CenterUniversity of CaliforniaDavisUSA
  3. 3.Department of Molecular Biosciences and BioengineeringUniversity of Hawaii at ManoaHonoluluUSA

Personalised recommendations