Establishment and Optimization of Monoclonal Antibody-based Heterologous dcELISA for 19-Nortestosterone Residue in Bovine Edible Tissue
Article first published online: 19 APR 2012
© 2012 Institute of Food Technologists®
Journal of Food Science
Volume 77, Issue 4, pages T63–T69, April 2012
How to Cite
Jiang, J., Zhang, H., Li, G., Yang, X., Li, R., Wang, Z. and Wang, J. (2012), Establishment and Optimization of Monoclonal Antibody-based Heterologous dcELISA for 19-Nortestosterone Residue in Bovine Edible Tissue. Journal of Food Science, 77: T63–T69. doi: 10.1111/j.1750-3841.2011.02578.x
- Issue published online: 19 APR 2012
- Article first published online: 19 APR 2012
- MS 20101257 Submitted 11/04/2011, Accepted 10/15/2011.
- heterologous dcELISA;
- monoclonal antibody;
Abstract: This paper presents the generation of monoclonal antibodies (mAbs) with high specificity against 19-nortestosterone (NT) through cell fusion procedures, and the development of mAb-based heterologous direct competitive enzyme-linked immunoabsorbent assay (dcELISA) methods to detect NT residue using one of these hybridomas (clone 3B8-E6). Under optimal experimental conditions, this assay exhibited a working range of 0.004 to 19 ng/mL with IC50 and limit of detection values of 0.28 and 0.002 ng/mL, respectively, when it was run in 0.01M phosphate-buffered saline (pH 7.4). Except for minor cross-reactivity with β-boldenone (6.9%) and trenbolone (1.2%), other interference to the assay was negligible (<0.05%). No significant differences (P > 0.05) were found for IC50 values when the pH of the assay buffer ranged from 6 to 8 and phosphate ion concentration was less than 20 mM. The dcELISA can tolerate higher concentrations of methanol than other organic solvents tested. When applied to bovine sample, the correlation coefficients (R) of the dcELISA and GC-MS data were 0.9918 in muscle, 0.9834 in liver, and 0.9976 in kidney. Therefore, this assay has the potential to be incorporated into a quantitative monitoring program for the rapid screening of NT residue in food.