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Protein microarray: sensitive and effective immunodetection for drug residues
BACKGROUND: Veterinary drugs such as clenbuterol (CL) and sulfamethazine (SM(2)) are low molecular weight (<1000 Da) compounds, or haptens, that are difficult to develop immunoassays due to their low immunogenicity. In this study, we conjugated the drugs to ovalbumin to increase their immunogenic...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2010
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2827365/ https://www.ncbi.nlm.nih.gov/pubmed/20158905 http://dx.doi.org/10.1186/1472-6750-10-12 |
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author | Zhong, Li Zhang, Wei Zer, Cindy Ge, Kun Gao, Xu Kernstine, Kemp H |
author_facet | Zhong, Li Zhang, Wei Zer, Cindy Ge, Kun Gao, Xu Kernstine, Kemp H |
author_sort | Zhong, Li |
collection | PubMed |
description | BACKGROUND: Veterinary drugs such as clenbuterol (CL) and sulfamethazine (SM(2)) are low molecular weight (<1000 Da) compounds, or haptens, that are difficult to develop immunoassays due to their low immunogenicity. In this study, we conjugated the drugs to ovalbumin to increase their immunogenicity for antiserum production in rabbits and developed a protein microarray immunoassay for detection of clenbuterol and sulfamethazine. The sensitivity of this approach was then compared to traditional ELISA technique. RESULTS: The artificial antigens were spotted on microarray slides. Standard concentrations of the compounds were added to compete with the spotted antigens for binding to the antisera to determine the IC(50). Our microarray assay showed the IC(50 )were 39.6 ng/ml for CL and 48.8 ng/ml for SM(2), while the traditional competitive indirect-ELISA (ci-ELISA) showed the IC(50 )were 190.7 ng/ml for CL and 156.7 ng/ml for SM(2). We further validated the two methods with CL fortified chicken muscle tissues, and the protein microarray assay showed 90% recovery while the ci-ELISA had 76% recovery rate. When tested with CL-fed chicken muscle tissues, the protein microarray assay had higher sensitivity (0.9 ng/g) than the ci-ELISA (0.1 ng/g) for detection of CL residues. CONCLUSIONS: The protein microarrays showed 4.5 and 3.5 times lower IC(50 )than the ci-ELISA detection for CL and SM(2), respectively, suggesting that immunodetection of small molecules with protein microarray is a better approach than the traditional ELISA technique. |
format | Text |
id | pubmed-2827365 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-28273652010-02-24 Protein microarray: sensitive and effective immunodetection for drug residues Zhong, Li Zhang, Wei Zer, Cindy Ge, Kun Gao, Xu Kernstine, Kemp H BMC Biotechnol Research article BACKGROUND: Veterinary drugs such as clenbuterol (CL) and sulfamethazine (SM(2)) are low molecular weight (<1000 Da) compounds, or haptens, that are difficult to develop immunoassays due to their low immunogenicity. In this study, we conjugated the drugs to ovalbumin to increase their immunogenicity for antiserum production in rabbits and developed a protein microarray immunoassay for detection of clenbuterol and sulfamethazine. The sensitivity of this approach was then compared to traditional ELISA technique. RESULTS: The artificial antigens were spotted on microarray slides. Standard concentrations of the compounds were added to compete with the spotted antigens for binding to the antisera to determine the IC(50). Our microarray assay showed the IC(50 )were 39.6 ng/ml for CL and 48.8 ng/ml for SM(2), while the traditional competitive indirect-ELISA (ci-ELISA) showed the IC(50 )were 190.7 ng/ml for CL and 156.7 ng/ml for SM(2). We further validated the two methods with CL fortified chicken muscle tissues, and the protein microarray assay showed 90% recovery while the ci-ELISA had 76% recovery rate. When tested with CL-fed chicken muscle tissues, the protein microarray assay had higher sensitivity (0.9 ng/g) than the ci-ELISA (0.1 ng/g) for detection of CL residues. CONCLUSIONS: The protein microarrays showed 4.5 and 3.5 times lower IC(50 )than the ci-ELISA detection for CL and SM(2), respectively, suggesting that immunodetection of small molecules with protein microarray is a better approach than the traditional ELISA technique. BioMed Central 2010-02-16 /pmc/articles/PMC2827365/ /pubmed/20158905 http://dx.doi.org/10.1186/1472-6750-10-12 Text en Copyright ©2010 Zhong et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research article Zhong, Li Zhang, Wei Zer, Cindy Ge, Kun Gao, Xu Kernstine, Kemp H Protein microarray: sensitive and effective immunodetection for drug residues |
title | Protein microarray: sensitive and effective immunodetection for drug residues |
title_full | Protein microarray: sensitive and effective immunodetection for drug residues |
title_fullStr | Protein microarray: sensitive and effective immunodetection for drug residues |
title_full_unstemmed | Protein microarray: sensitive and effective immunodetection for drug residues |
title_short | Protein microarray: sensitive and effective immunodetection for drug residues |
title_sort | protein microarray: sensitive and effective immunodetection for drug residues |
topic | Research article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2827365/ https://www.ncbi.nlm.nih.gov/pubmed/20158905 http://dx.doi.org/10.1186/1472-6750-10-12 |
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