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Preparation of polyclonal antibody and development of a biotin-streptavidin-based ELISA method for detecting kanamycin in milk and honey

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Abstract

Kanamycin is an aminoglycoside antibiotic used increasingly in human and veterinary medicine. However, kanamycin residues in food can cause serious side effects. Here we reported the preparation of polyclonal antibody and the development of an indirect competitive biotin-streptavidin-amplified-based enzyme-linked immunosorbent assay(BA-ELISA) that can sensitively and specifically detect kanamycin residues in milk and honey. The immunogen and coating antigen were synthesized by covalently linking kanamycin to carrier proteins using the carbodiimide method. The anti-kanamycin polyclonal antibodies were obtained from immunized rabbits. The key assay parameters were investigated and optimized. The results show that under optimum conditions, the limit of detection for kanamycin is 0.07 ng/mL and the IC50 is 6.48 ng/mL. Cross-reactivity values of the antibody with four kanamycin analogues are all <1%. Trace amounts of kanamycin in milk and honey samples can be detected by this novel BA-ELISA method successfully with satisfactory recoveries of 91.0%—103.3%. The developed protocol was also validated against liquid chromatography-mass spectrometry, returning a significant correlation. These results indicate that BA-ELISA is a viable option for monitoring kanamycin residues in milk and honey.

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References

  1. Magnet S., Blanchard J. S., Chem. Rev., 2005, 105, 477

    Article  CAS  PubMed  Google Scholar 

  2. Joshi T., Voo Z. X., Graham B., Spiccia L., Martin L. L., Biochim. Biophys. Acta, 2015, 1848, 385

    Article  CAS  PubMed  Google Scholar 

  3. Ramezani M., Danesh N. M., Lavaee P., Abnous K., Taghdisi S. M., Sensors Actua. B: Chem., 2016, 222, 1

    Article  CAS  Google Scholar 

  4. Luft F. C., Bloch R., Sloan R. S., Yum M. N., Costello R., Maxwell D. R., J. Infec. Dis., 1978, 138, 541

    Article  CAS  Google Scholar 

  5. EMEA/MRL/886/03-FINAL, 2003, http://www.emea.europa.eu/pdfs/ vet/mrls/Mrlopinions/094203en.pdf.

  6. Preua M., Guyot G., Petza M., J. Chromatogr. A, 1998, 818, 95

    Article  Google Scholar 

  7. Chen S., Liang Y., Chou Y., J. Sep. Sci., 2006, 29, 607

    Article  CAS  PubMed  Google Scholar 

  8. Oertel R., Neumeister V., Kirch W., J. Chromatogr. A, 2004, 1058, 197

    Article  CAS  PubMed  Google Scholar 

  9. Tao Y., Chen D., Yu H., Huang L., Liu Z., Cao X., Yan C., Pan Y., Liu Z., Yuan Z., Food Chem., 2012, 135, 676

    Article  CAS  PubMed  Google Scholar 

  10. Longa Y. H., Hernandezb M., Kaalea E., Schepdael A. V., Roetsa E., Borrull F., Calull M., Hoogmartens J., J. Chromatogr. B, 2003, 784, 255

    Article  Google Scholar 

  11. Kaale E., Schepdael A. V., Roets E., Hoogmartens J., Electrophoresis, 2003, 24, 1119

    Article  CAS  PubMed  Google Scholar 

  12. Watanabe H., Satake A., Kido Y., Tsuji A., Analyst, 1999, 124, 1611

    Article  CAS  PubMed  Google Scholar 

  13. Loomans E. E. M. G., Wiltenburg J. V., Koets M., Amerongen A. V., J. Agric. Food Chem., 2003, 51, 587

    Article  CAS  PubMed  Google Scholar 

  14. Chen Y., Wang Z., Wang Z., Tang S., Zhu Y., **ao X., J. Agric. Food Chem., 2008, 56, 2944

    Article  CAS  PubMed  Google Scholar 

  15. Zhu T., Row K., Chem. Res. Chinese Universities, 2013, 29(3), 429

    Article  CAS  Google Scholar 

  16. Samdal I. A., Lovberg K. E., Briggs L. R., Kilcoyne J., Xu J., Forsyth C. J., Miles C. O., J. Agric. Food Chem., 2015, 63, 7855

    Article  CAS  PubMed  Google Scholar 

  17. Yang F., Zhu L., Meng W., Liu R., Anal. Methods, 2015, 7, 5275

    Article  CAS  Google Scholar 

  18. Leivo J., Makela J., Rosenberg J., Lamminmaki U., Anal. Biochem., 2016, 492, 27

    Article  CAS  PubMed  Google Scholar 

  19. Liang Y., Huang X., Yu R., Zhou Y., **ong Y., Anal. Chim. Acta, 2016, 936, 195

    Article  CAS  PubMed  Google Scholar 

  20. Zhan S., Huang X., Chen R., Li J., **ong Y., Talanta, 2016, 158, 51

    Article  CAS  PubMed  Google Scholar 

  21. Novo P., Moulas G., Prazeres D. M. F., Chu V., Conde J. P., Sensors Actua. B: Chem., 2013, 176, 232

    Article  CAS  Google Scholar 

  22. Liu B., Ge Y., Zhang Y., Song Y., Chen Y., Wang S., Anal. Methods, 2013, 5, 5938

    Article  CAS  Google Scholar 

  23. Wang S., Ge L., Song X., Yu J., Ge S., Huang J., Zeng F., Biosens. Bioelectron., 2012, 31, 212

    Article  CAS  PubMed  Google Scholar 

  24. Liu N., Nie D., Zhao Z., Meng X., Wu A., Food Control, 2015, 57, 202

    Article  CAS  Google Scholar 

  25. Bu D., Zhuang H., Zhou X., Yang G., Anal. Biochem., 2014, 462, 51

    Article  CAS  PubMed  Google Scholar 

  26. Fang X. X., Zheng J., Yan G. Q., Chem. Res. Chinese Universities, 2016, 32(6), 917

    Article  CAS  Google Scholar 

  27. Harada H., Onoda A., Uematsu T., Kuwabata S., Hayashi T., Lang-muir, 2016, 32, 6459

    Article  CAS  Google Scholar 

  28. Zhang Y., Gao X., Gao A., Fan M., Food Anal. Methods, 2012, 5, 1214

    Article  Google Scholar 

  29. Du H., Chu Y., Yang H., Zhao K., Li J., She P., Zhang X., Deng A., Anal. Methods, 2016, 8, 3578

    Article  CAS  Google Scholar 

  30. Bai Y., Liu Z., Bi Y., Wang X., ** Y., Sun L., Wang H., Zhang C., Xu S., J. Agric. Food Chem., 2012, 60, 11618

    Article  CAS  PubMed  Google Scholar 

  31. GB/T 22969-2008, Determination of Streptomycin, Dihydrostrepto-mycin and Kanamycin Residues in Milk and Milk Powder-LC- MS-MS Method

  32. GB/T 22995-2008, Determination of Streptomycin, Dihydrostrepto-mycin and Kanamycin Residues in Honey-LC-MS-MS Method

  33. Jiang W., Beier R. C., Luo P., Zhai P., Wu N., Lin G., Wang X., Xu G., J. Agric. Food Chem., 2016, 64, 364

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Bei**g Key Laboratory of Environmentally Harmful Chemical Analysis, College of Science, Bei**g University of Chemical Technology, Bei**g, 100029, P. R. China

    ** Su, **aonan Chen, Zhang**g He & Yi Yang

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  1. ** Su
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  2. **aonan Chen
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  3. Zhang**g He
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  4. Yi Yang
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Corresponding author

Correspondence to Yi Yang.

Additional information

Supported by the National Natural Science Foundation of China(No.21675008), the Natural Science Foundation of Bei**g, China(No.2132048) and the Fundamental Research Funds for the Central Universities, China(No.JD1516).

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40242_2017_7168_MOESM1_ESM.pdf

Preparation of polyclonal antibody and development of a biotin-streptavidin-based ELISA method for detecting kanamycin in milk and honey

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Su, P., Chen, X., He, Z. et al. Preparation of polyclonal antibody and development of a biotin-streptavidin-based ELISA method for detecting kanamycin in milk and honey. Chem. Res. Chin. Univ. 33, 876–881 (2017). https://doi.org/10.1007/s40242-017-7168-9

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  • DOI: https://doi.org/10.1007/s40242-017-7168-9

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