Document Detail


Monoclonal Antibody-based ELISA and Colloidal Gold Immunoassay for Detecting 19-Nortestosterone Residue in Animal Tissues.
MedLine Citation:
PMID:  21854067     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
This paper presents the generation of monoclonal antibodies (mAbs) with high specificity against 19-nortestosterone (NT) through cell fusion techniques, and the development of a mAb-based indirect competitive ELISA (icELISA) method and colloidal gold-based immuno-chromatographic assay to detect NT residue in beef and pork samples. Modified carbodiimide method was employed to synthesize the artificial antigen and BALB/c mice were used to produce anti-NT mAbs. Based on the checkerboard titration, an indirect competitive ELISA standard curve was established. This assay was sensitive and had a linear range from 0.03 to 38 ng/mL in phosphate buffered saline (PBS), with IC50 and LOD values of 0.52 ng/mL and 0.01 ng/mL, respectively. Of all the competitive analogues, the produced mAb exhibited a high cross-reactivity to 17α-nortestosterone (83.6%), the main metabolite of NT in animal tissues. Except for moderate cross-reactivities with trenbolone (22.6%) and β-boldenone (13.8%), the other interference to the assay was negligible (<0.05%). In contrast, the strip test had a visual detection limit of 1 ng/mL in PBS, 2 μg/kg in beef, and 2 μg/kg in pork, respectively; and the results can be judged within 10 min. The ELISA and GC-MS results showed close correlation in beef (R2= 0.9945) and in pork (R2= 0.9977). Therefore, the combination of two immunoassays provides a useful screening method for quantitative, or qualitative detection of NT residue in animal-original products.
Authors:
Jinqing Jiang; Ziliang Wang; Haitang Zhang; Xiaojian Zhang; Xingyou Liu; Sanhu Wang
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-8-22
Journal Detail:
Title:  Journal of agricultural and food chemistry     Volume:  -     ISSN:  1520-5118     ISO Abbreviation:  -     Publication Date:  2011 Aug 
Date Detail:
Created Date:  2011-8-22     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0374755     Medline TA:  J Agric Food Chem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
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