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Enhanced Sensitive Immunoassay: Noncompetitive Phage Anti-Immune Complex Assay for the Determination of Malachite Green and Leucomalachite Green
[Image: see text] To develop a more sensitive immunoassay for malachite green (MG) and leucomalachite green (LMG), we identified the immunocomplex binding phage-borne peptides for use in the noncompetitive phage anti-immunocomplex assay (PHAIA). An anti-LMG monoclonal antibody (mAb) was used to sele...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical
Society
2014
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4150606/ https://www.ncbi.nlm.nih.gov/pubmed/25077381 http://dx.doi.org/10.1021/jf5019824 |
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author | Dong, Jie-Xian Xu, Chao Wang, Hong Xiao, Zhi-Li Gee, Shirley J. Li, Zhen-Feng Wang, Feng Wu, Wei-Jian Shen, Yu-Dong Yang, Jin-Yi Sun, Yuan-Ming Hammock, Bruce D. |
author_facet | Dong, Jie-Xian Xu, Chao Wang, Hong Xiao, Zhi-Li Gee, Shirley J. Li, Zhen-Feng Wang, Feng Wu, Wei-Jian Shen, Yu-Dong Yang, Jin-Yi Sun, Yuan-Ming Hammock, Bruce D. |
author_sort | Dong, Jie-Xian |
collection | PubMed |
description | [Image: see text] To develop a more sensitive immunoassay for malachite green (MG) and leucomalachite green (LMG), we identified the immunocomplex binding phage-borne peptides for use in the noncompetitive phage anti-immunocomplex assay (PHAIA). An anti-LMG monoclonal antibody (mAb) was used to select immunocomplex binding peptides from a circular random eight-amino-acid phage-displayed library. After three rounds of panning-elution, five peptides that bound the LMG–mAb immunocomplex were obtained. One of the phage-borne peptide clones that resulted in an assay with the highest sensitivity was chosen for further research. The concentration of LMG producing 50% of the saturated signal and the limit of detection of the assay were 7.02 and 0.55 ng/mL, respectively, with a linear range of 1.35 to 21.56 ng/mL. The PHAIA based on the same antibody was 16 times more sensitive compared to the competitive immunoassay. PHAIA was used to analyze LMG, MG, and two mixtures of spiked fish samples, with validation by high-performance liquid chromatography (HPLC) with fluorescence detector. Results showed a good correlation (R(2)(LMG) = 0.9841; R(2)(MG) = 0.993; R(2)(Mixture) = 0.9903) between the data of PHAIA and HPLC, thus the assay was an efficient method for monitoring food safety. |
format | Online Article Text |
id | pubmed-4150606 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | American Chemical
Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-41506062015-07-31 Enhanced Sensitive Immunoassay: Noncompetitive Phage Anti-Immune Complex Assay for the Determination of Malachite Green and Leucomalachite Green Dong, Jie-Xian Xu, Chao Wang, Hong Xiao, Zhi-Li Gee, Shirley J. Li, Zhen-Feng Wang, Feng Wu, Wei-Jian Shen, Yu-Dong Yang, Jin-Yi Sun, Yuan-Ming Hammock, Bruce D. J Agric Food Chem [Image: see text] To develop a more sensitive immunoassay for malachite green (MG) and leucomalachite green (LMG), we identified the immunocomplex binding phage-borne peptides for use in the noncompetitive phage anti-immunocomplex assay (PHAIA). An anti-LMG monoclonal antibody (mAb) was used to select immunocomplex binding peptides from a circular random eight-amino-acid phage-displayed library. After three rounds of panning-elution, five peptides that bound the LMG–mAb immunocomplex were obtained. One of the phage-borne peptide clones that resulted in an assay with the highest sensitivity was chosen for further research. The concentration of LMG producing 50% of the saturated signal and the limit of detection of the assay were 7.02 and 0.55 ng/mL, respectively, with a linear range of 1.35 to 21.56 ng/mL. The PHAIA based on the same antibody was 16 times more sensitive compared to the competitive immunoassay. PHAIA was used to analyze LMG, MG, and two mixtures of spiked fish samples, with validation by high-performance liquid chromatography (HPLC) with fluorescence detector. Results showed a good correlation (R(2)(LMG) = 0.9841; R(2)(MG) = 0.993; R(2)(Mixture) = 0.9903) between the data of PHAIA and HPLC, thus the assay was an efficient method for monitoring food safety. American Chemical Society 2014-07-31 2014-08-27 /pmc/articles/PMC4150606/ /pubmed/25077381 http://dx.doi.org/10.1021/jf5019824 Text en Copyright © 2014 American Chemical Society Terms of Use (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) |
spellingShingle | Dong, Jie-Xian Xu, Chao Wang, Hong Xiao, Zhi-Li Gee, Shirley J. Li, Zhen-Feng Wang, Feng Wu, Wei-Jian Shen, Yu-Dong Yang, Jin-Yi Sun, Yuan-Ming Hammock, Bruce D. Enhanced Sensitive Immunoassay: Noncompetitive Phage Anti-Immune Complex Assay for the Determination of Malachite Green and Leucomalachite Green |
title | Enhanced
Sensitive Immunoassay: Noncompetitive Phage
Anti-Immune Complex Assay for the Determination of Malachite Green
and Leucomalachite Green |
title_full | Enhanced
Sensitive Immunoassay: Noncompetitive Phage
Anti-Immune Complex Assay for the Determination of Malachite Green
and Leucomalachite Green |
title_fullStr | Enhanced
Sensitive Immunoassay: Noncompetitive Phage
Anti-Immune Complex Assay for the Determination of Malachite Green
and Leucomalachite Green |
title_full_unstemmed | Enhanced
Sensitive Immunoassay: Noncompetitive Phage
Anti-Immune Complex Assay for the Determination of Malachite Green
and Leucomalachite Green |
title_short | Enhanced
Sensitive Immunoassay: Noncompetitive Phage
Anti-Immune Complex Assay for the Determination of Malachite Green
and Leucomalachite Green |
title_sort | enhanced
sensitive immunoassay: noncompetitive phage
anti-immune complex assay for the determination of malachite green
and leucomalachite green |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4150606/ https://www.ncbi.nlm.nih.gov/pubmed/25077381 http://dx.doi.org/10.1021/jf5019824 |
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