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Heterologous screening of hybridomas for the development of broad-specific monoclonal antibodies against deoxynivalenol and its analogues

Yirong Guo (UGent) , Melanie Sanders (UGent) , Anastasia Galvita (UGent) , Arne Heyerick (UGent) , Dieter Deforce (UGent) , Marc Bracke (UGent) , S Eremin and Sarah De Saeger (UGent)
(2014) WORLD MYCOTOXIN JOURNAL. 7(3). p.257-265
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Abstract
Hapten heterology was introduced into the steps of hybridoma selection for the development of monoclonal antibodies (MAbs) against deoxynivalenol (DON). Firstly, a novel heterologous DON hapten was synthesised and covalently coupled to proteins (i.e. bovine serum albumin (BSA), ovalbumin and horseradish peroxidase) using the linkage of cyanuric chloride (CC). After immunisation, antisera from different DON immunogens were checked for the presence of useful antibodies. Next, both homologous and heterologous enzyme-linked immunosorbent assays were conducted to screen for hybridomas. It was found that heterologous screening could significantly reduce the proportion of false positives and appeared to be an efficient approach for selecting hybridomas of interest. This strategy resulted in two kinds of broad-selective MAbs against DON and its analogues. They were quite distinct from other reported DON-antibodies in their cross-reactivity profiles. A unique MAb 13H1 derived from DON-CC-BSA immunogen could recognise DON and its analogues in the order of HT-2 toxin > 15-acetyl-DON > DON > nivalenol, with IC50 ranging from 1.14 to 7.69 mu g/ml. Another preferable MAb 10H10 generated from DON-BSA immunogen manifested relatively similar affinity to DON, 3-acetyl-DON and 15-acetyl-DON, with IC50 values of 22, 15 and 34 ng/ml, respectively. This is the first broad-specific MAb against DON and its two acetylated forms and thus it can be used for simultaneous detection of the three mycotoxins.
Keywords
IMMUNOASSAY, HAPTEN, MOLECULAR-WEIGHT, CROSS-REACTIVITY, LINKED-IMMUNOSORBENT-ASSAY, MYCOTOXINS, PROTEINS, ELISA, WHEAT, hybridomas, hapten heterology, cross-reactivity, competitive ELISA, monoclonal antibody, deoxynivalenol

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Citation

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MLA
Guo, Yirong et al. “Heterologous Screening of Hybridomas for the Development of Broad-specific Monoclonal Antibodies Against Deoxynivalenol and Its Analogues.” WORLD MYCOTOXIN JOURNAL 7.3 (2014): 257–265. Print.
APA
Guo, Yirong, Sanders, M., Galvita, A., Heyerick, A., Deforce, D., Bracke, M., Eremin, S., et al. (2014). Heterologous screening of hybridomas for the development of broad-specific monoclonal antibodies against deoxynivalenol and its analogues. WORLD MYCOTOXIN JOURNAL, 7(3), 257–265.
Chicago author-date
Guo, Yirong, Melanie Sanders, Anastasia Galvita, Arne Heyerick, Dieter Deforce, Marc Bracke, S Eremin, and Sarah De Saeger. 2014. “Heterologous Screening of Hybridomas for the Development of Broad-specific Monoclonal Antibodies Against Deoxynivalenol and Its Analogues.” World Mycotoxin Journal 7 (3): 257–265.
Chicago author-date (all authors)
Guo, Yirong, Melanie Sanders, Anastasia Galvita, Arne Heyerick, Dieter Deforce, Marc Bracke, S Eremin, and Sarah De Saeger. 2014. “Heterologous Screening of Hybridomas for the Development of Broad-specific Monoclonal Antibodies Against Deoxynivalenol and Its Analogues.” World Mycotoxin Journal 7 (3): 257–265.
Vancouver
1.
Guo Y, Sanders M, Galvita A, Heyerick A, Deforce D, Bracke M, et al. Heterologous screening of hybridomas for the development of broad-specific monoclonal antibodies against deoxynivalenol and its analogues. WORLD MYCOTOXIN JOURNAL. 2014;7(3):257–65.
IEEE
[1]
Y. Guo et al., “Heterologous screening of hybridomas for the development of broad-specific monoclonal antibodies against deoxynivalenol and its analogues,” WORLD MYCOTOXIN JOURNAL, vol. 7, no. 3, pp. 257–265, 2014.
@article{5766859,
  abstract     = {Hapten heterology was introduced into the steps of hybridoma selection for the development of monoclonal antibodies (MAbs) against deoxynivalenol (DON). Firstly, a novel heterologous DON hapten was synthesised and covalently coupled to proteins (i.e. bovine serum albumin (BSA), ovalbumin and horseradish peroxidase) using the linkage of cyanuric chloride (CC). After immunisation, antisera from different DON immunogens were checked for the presence of useful antibodies. Next, both homologous and heterologous enzyme-linked immunosorbent assays were conducted to screen for hybridomas. It was found that heterologous screening could significantly reduce the proportion of false positives and appeared to be an efficient approach for selecting hybridomas of interest. This strategy resulted in two kinds of broad-selective MAbs against DON and its analogues. They were quite distinct from other reported DON-antibodies in their cross-reactivity profiles. A unique MAb 13H1 derived from DON-CC-BSA immunogen could recognise DON and its analogues in the order of HT-2 toxin > 15-acetyl-DON > DON > nivalenol, with IC50 ranging from 1.14 to 7.69 mu g/ml. Another preferable MAb 10H10 generated from DON-BSA immunogen manifested relatively similar affinity to DON, 3-acetyl-DON and 15-acetyl-DON, with IC50 values of 22, 15 and 34 ng/ml, respectively. This is the first broad-specific MAb against DON and its two acetylated forms and thus it can be used for simultaneous detection of the three mycotoxins.},
  author       = {Guo, Yirong and Sanders, Melanie and Galvita, Anastasia and Heyerick, Arne and Deforce, Dieter and Bracke, Marc and Eremin, S and De Saeger, Sarah},
  issn         = {1875-0710},
  journal      = {WORLD MYCOTOXIN JOURNAL},
  keywords     = {IMMUNOASSAY,HAPTEN,MOLECULAR-WEIGHT,CROSS-REACTIVITY,LINKED-IMMUNOSORBENT-ASSAY,MYCOTOXINS,PROTEINS,ELISA,WHEAT,hybridomas,hapten heterology,cross-reactivity,competitive ELISA,monoclonal antibody,deoxynivalenol},
  language     = {eng},
  number       = {3},
  pages        = {257--265},
  title        = {Heterologous screening of hybridomas for the development of broad-specific monoclonal antibodies against deoxynivalenol and its analogues},
  url          = {http://dx.doi.org/10.3920/WMJ2013.1668},
  volume       = {7},
  year         = {2014},
}

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