Highly Sensitive Immunochromatographic Identification of Tetracycline Antibiotics in Milk

A rapid immunochromatographic assay was developed for the control of tetracycline (TC). The assay is based on the competition between immobilized TC-protein conjugate and TC in a tested sample for binding with polyclonal anti-TC antibodies conjugated to colloidal gold during the flow of the sample a...

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Main Authors: N. A. Taranova, A. S. Kruhlik, E. A. Zvereva, V. V. Shmanai, I. I. Vashkevich, D. A. Semyonov, S. A. Eremin, A. V. Zherdev, B. B. Dzantiev
Format: Article
Language:English
Published: Wiley 2015-01-01
Series:International Journal of Analytical Chemistry
Online Access:http://dx.doi.org/10.1155/2015/347621
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author N. A. Taranova
A. S. Kruhlik
E. A. Zvereva
V. V. Shmanai
I. I. Vashkevich
D. A. Semyonov
S. A. Eremin
A. V. Zherdev
B. B. Dzantiev
author_facet N. A. Taranova
A. S. Kruhlik
E. A. Zvereva
V. V. Shmanai
I. I. Vashkevich
D. A. Semyonov
S. A. Eremin
A. V. Zherdev
B. B. Dzantiev
author_sort N. A. Taranova
collection DOAJ
description A rapid immunochromatographic assay was developed for the control of tetracycline (TC). The assay is based on the competition between immobilized TC-protein conjugate and TC in a tested sample for binding with polyclonal anti-TC antibodies conjugated to colloidal gold during the flow of the sample along a membrane strip with immobilized reactants. Conjugation of colloidal gold and the total immunoglobulin (IgG) fraction of polyclonal antibodies was used to increase the assay sensitivity to ensure low content of specific antibodies in the conjugate. This allowed effective inhibition of free TC and conjugate binding in the strip test zone. Photometric marker registration allows control of the reduction of binding, thereby enhancing detection sensitivity. The proposed assay allows TC to be detected at concentrations up to 20 ng/mL, exceeding the limit of detection of the known analogues, in a wide working range (more than two orders) of 60 pg/mL to 10 ng/mL, ensured through the use of polyclonal antibodies. The assay time is 10 min. The efficiency of the designed assay is shown to identify TC in milk; the degree of recovery of TC ranges from 90 to 112%. The precision of the concentrations measurements was no more than 10%.
format Article
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institution Kabale University
issn 1687-8760
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publishDate 2015-01-01
publisher Wiley
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series International Journal of Analytical Chemistry
spelling doaj-art-bb16b8b378f14375ac37454dc3d0a94f2025-02-03T06:05:31ZengWileyInternational Journal of Analytical Chemistry1687-87601687-87792015-01-01201510.1155/2015/347621347621Highly Sensitive Immunochromatographic Identification of Tetracycline Antibiotics in MilkN. A. Taranova0A. S. Kruhlik1E. A. Zvereva2V. V. Shmanai3I. I. Vashkevich4D. A. Semyonov5S. A. Eremin6A. V. Zherdev7B. B. Dzantiev8A.N. Bach Institute of Biochemistry, Research Centre of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, Moscow 119071, RussiaInstitute of Physical Organic Chemistry, Surganov Street 13, 220072 Minsk, BelarusA.N. Bach Institute of Biochemistry, Research Centre of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, Moscow 119071, RussiaInstitute of Physical Organic Chemistry, Surganov Street 13, 220072 Minsk, BelarusInstitute of Bioorganic Chemistry, Acad. Kuprevich Street 5/2, 220141 Minsk, BelarusInstitute of Bioorganic Chemistry, Acad. Kuprevich Street 5/2, 220141 Minsk, BelarusA.N. Bach Institute of Biochemistry, Research Centre of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, Moscow 119071, RussiaA.N. Bach Institute of Biochemistry, Research Centre of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, Moscow 119071, RussiaA.N. Bach Institute of Biochemistry, Research Centre of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, Moscow 119071, RussiaA rapid immunochromatographic assay was developed for the control of tetracycline (TC). The assay is based on the competition between immobilized TC-protein conjugate and TC in a tested sample for binding with polyclonal anti-TC antibodies conjugated to colloidal gold during the flow of the sample along a membrane strip with immobilized reactants. Conjugation of colloidal gold and the total immunoglobulin (IgG) fraction of polyclonal antibodies was used to increase the assay sensitivity to ensure low content of specific antibodies in the conjugate. This allowed effective inhibition of free TC and conjugate binding in the strip test zone. Photometric marker registration allows control of the reduction of binding, thereby enhancing detection sensitivity. The proposed assay allows TC to be detected at concentrations up to 20 ng/mL, exceeding the limit of detection of the known analogues, in a wide working range (more than two orders) of 60 pg/mL to 10 ng/mL, ensured through the use of polyclonal antibodies. The assay time is 10 min. The efficiency of the designed assay is shown to identify TC in milk; the degree of recovery of TC ranges from 90 to 112%. The precision of the concentrations measurements was no more than 10%.http://dx.doi.org/10.1155/2015/347621
spellingShingle N. A. Taranova
A. S. Kruhlik
E. A. Zvereva
V. V. Shmanai
I. I. Vashkevich
D. A. Semyonov
S. A. Eremin
A. V. Zherdev
B. B. Dzantiev
Highly Sensitive Immunochromatographic Identification of Tetracycline Antibiotics in Milk
International Journal of Analytical Chemistry
title Highly Sensitive Immunochromatographic Identification of Tetracycline Antibiotics in Milk
title_full Highly Sensitive Immunochromatographic Identification of Tetracycline Antibiotics in Milk
title_fullStr Highly Sensitive Immunochromatographic Identification of Tetracycline Antibiotics in Milk
title_full_unstemmed Highly Sensitive Immunochromatographic Identification of Tetracycline Antibiotics in Milk
title_short Highly Sensitive Immunochromatographic Identification of Tetracycline Antibiotics in Milk
title_sort highly sensitive immunochromatographic identification of tetracycline antibiotics in milk
url http://dx.doi.org/10.1155/2015/347621
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