Monoclonal antibody enzyme‐linked immunosorbent assay to quantify vitellogenin for studies on environmental estrogens in the rainbow trout ( Oncorhynchus mykiss)
Abstract Vitellogenin (VTG) induction has proved to be a valuable biomarker for assessing exposure to environmental estrogens in fish. The widespread use of VTG in this regard has lead to the need for standardized assays to quantify VTG, and monoclonal antibodies have the potential to help accomplis...
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crwiley:10.1002/etc.5620210107 2024-06-02T08:03:43+00:00 Monoclonal antibody enzyme‐linked immunosorbent assay to quantify vitellogenin for studies on environmental estrogens in the rainbow trout ( Oncorhynchus mykiss) Tyler, Charles R. van Aerle, Ronny Nilsen, Mona V. Blackwell, Robert Maddix, Susan Nilsen, Bente M. Berg, Karin Hutchinson, Tom H. Goksøyr, Anders 2002 http://dx.doi.org/10.1002/etc.5620210107 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fetc.5620210107 https://setac.onlinelibrary.wiley.com/doi/pdf/10.1002/etc.5620210107 en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor Environmental Toxicology and Chemistry volume 21, issue 1, page 47-54 ISSN 0730-7268 1552-8618 journal-article 2002 crwiley https://doi.org/10.1002/etc.5620210107 2024-05-03T11:38:16Z Abstract Vitellogenin (VTG) induction has proved to be a valuable biomarker for assessing exposure to environmental estrogens in fish. The widespread use of VTG in this regard has lead to the need for standardized assays to quantify VTG, and monoclonal antibodies have the potential to help accomplish this. A VTG enzyme‐linked immunosorbent assay (ELISA) was developed using a monoclonal antibody prepared against Atlantic salmon ( Salmo salar ) VTG (MAb BN‐5) and its ability to quantify VTG in the rainbow trout ( Oncorhynchus mykiss ) compared with a rainbow trout vitellogenin (rt‐VTG) ELISA that employed homologous polyclonal antibodies (PAb). In routine protocols, the working range of the homologous rt‐PAb VTG ELISA was between 9 ng/ml and 70 ng/ml (80–20% relative maximum binding [B/Bo]) with a 50% B/Bo of 25 ± 0.9 ng/ml and inter‐ and intraassay variations at 50% B/Bo of 7% ( n = 7) and 8% ( n = 15), respectively. The working range of the MAb BN‐5 VTG ELISA was between 60 ng/ml and 850 ng/ml (80–20% B/Bo) with a 50% B/Bo of 227 ± 22 ng/ml and inter‐ and intraassay variations at 50% B/Bo of 5% ( n = 10) and 9% ( n = 12), respectively. In the routine protocols, detection limits for measurement of plasma VTG in rainbow trout (at 80% B/Bo; and given the requirement to dilute plasma to a minimum of 1:10 for the assays) were 90 ng/ml for the polyclonal rt‐VTG assay and approximately 600 ng/ml in the monoclonal antibody assay. In juvenile female rainbow trout exposed to a series of doses of estradiol‐17β (E2) and 4‐ tert nonylphenol (4‐NP), there were no differences in the vitellogenic responses measured in the PAb and MAb BN‐5 VTG ELISAs. The monoclonal MAb BN‐5 VTG ELISA is likely to be of considerable value for studies on environmental estrogens in juvenile female rainbow trout in standardized tests. Article in Journal/Newspaper Atlantic salmon Salmo salar Wiley Online Library Environmental Toxicology and Chemistry 21 1 47 54 |
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Wiley Online Library |
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English |
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Abstract Vitellogenin (VTG) induction has proved to be a valuable biomarker for assessing exposure to environmental estrogens in fish. The widespread use of VTG in this regard has lead to the need for standardized assays to quantify VTG, and monoclonal antibodies have the potential to help accomplish this. A VTG enzyme‐linked immunosorbent assay (ELISA) was developed using a monoclonal antibody prepared against Atlantic salmon ( Salmo salar ) VTG (MAb BN‐5) and its ability to quantify VTG in the rainbow trout ( Oncorhynchus mykiss ) compared with a rainbow trout vitellogenin (rt‐VTG) ELISA that employed homologous polyclonal antibodies (PAb). In routine protocols, the working range of the homologous rt‐PAb VTG ELISA was between 9 ng/ml and 70 ng/ml (80–20% relative maximum binding [B/Bo]) with a 50% B/Bo of 25 ± 0.9 ng/ml and inter‐ and intraassay variations at 50% B/Bo of 7% ( n = 7) and 8% ( n = 15), respectively. The working range of the MAb BN‐5 VTG ELISA was between 60 ng/ml and 850 ng/ml (80–20% B/Bo) with a 50% B/Bo of 227 ± 22 ng/ml and inter‐ and intraassay variations at 50% B/Bo of 5% ( n = 10) and 9% ( n = 12), respectively. In the routine protocols, detection limits for measurement of plasma VTG in rainbow trout (at 80% B/Bo; and given the requirement to dilute plasma to a minimum of 1:10 for the assays) were 90 ng/ml for the polyclonal rt‐VTG assay and approximately 600 ng/ml in the monoclonal antibody assay. In juvenile female rainbow trout exposed to a series of doses of estradiol‐17β (E2) and 4‐ tert nonylphenol (4‐NP), there were no differences in the vitellogenic responses measured in the PAb and MAb BN‐5 VTG ELISAs. The monoclonal MAb BN‐5 VTG ELISA is likely to be of considerable value for studies on environmental estrogens in juvenile female rainbow trout in standardized tests. |
format |
Article in Journal/Newspaper |
author |
Tyler, Charles R. van Aerle, Ronny Nilsen, Mona V. Blackwell, Robert Maddix, Susan Nilsen, Bente M. Berg, Karin Hutchinson, Tom H. Goksøyr, Anders |
spellingShingle |
Tyler, Charles R. van Aerle, Ronny Nilsen, Mona V. Blackwell, Robert Maddix, Susan Nilsen, Bente M. Berg, Karin Hutchinson, Tom H. Goksøyr, Anders Monoclonal antibody enzyme‐linked immunosorbent assay to quantify vitellogenin for studies on environmental estrogens in the rainbow trout ( Oncorhynchus mykiss) |
author_facet |
Tyler, Charles R. van Aerle, Ronny Nilsen, Mona V. Blackwell, Robert Maddix, Susan Nilsen, Bente M. Berg, Karin Hutchinson, Tom H. Goksøyr, Anders |
author_sort |
Tyler, Charles R. |
title |
Monoclonal antibody enzyme‐linked immunosorbent assay to quantify vitellogenin for studies on environmental estrogens in the rainbow trout ( Oncorhynchus mykiss) |
title_short |
Monoclonal antibody enzyme‐linked immunosorbent assay to quantify vitellogenin for studies on environmental estrogens in the rainbow trout ( Oncorhynchus mykiss) |
title_full |
Monoclonal antibody enzyme‐linked immunosorbent assay to quantify vitellogenin for studies on environmental estrogens in the rainbow trout ( Oncorhynchus mykiss) |
title_fullStr |
Monoclonal antibody enzyme‐linked immunosorbent assay to quantify vitellogenin for studies on environmental estrogens in the rainbow trout ( Oncorhynchus mykiss) |
title_full_unstemmed |
Monoclonal antibody enzyme‐linked immunosorbent assay to quantify vitellogenin for studies on environmental estrogens in the rainbow trout ( Oncorhynchus mykiss) |
title_sort |
monoclonal antibody enzyme‐linked immunosorbent assay to quantify vitellogenin for studies on environmental estrogens in the rainbow trout ( oncorhynchus mykiss) |
publisher |
Wiley |
publishDate |
2002 |
url |
http://dx.doi.org/10.1002/etc.5620210107 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fetc.5620210107 https://setac.onlinelibrary.wiley.com/doi/pdf/10.1002/etc.5620210107 |
genre |
Atlantic salmon Salmo salar |
genre_facet |
Atlantic salmon Salmo salar |
op_source |
Environmental Toxicology and Chemistry volume 21, issue 1, page 47-54 ISSN 0730-7268 1552-8618 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1002/etc.5620210107 |
container_title |
Environmental Toxicology and Chemistry |
container_volume |
21 |
container_issue |
1 |
container_start_page |
47 |
op_container_end_page |
54 |
_version_ |
1800748333878214656 |