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Development of anti-immunocomplex specific antibodies and non-competitive time-resolved fluorescence immunoassay for the detection of estradiol

Detection of circulatory estradiol has widespread use in various clinical applications. Particularly, the use of estradiol-specific antibodies in immunoassays is routinely used, mainly due to the cost efficiency and simplicity of the sample handling process. However, the circulatory levels of estrad...

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Autores principales: Leivo, Janne, Kivimäki, Laura, Juntunen, Etvi, Pettersson, Kim, Lamminmäki, Urpo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6704259/
https://www.ncbi.nlm.nih.gov/pubmed/31177333
http://dx.doi.org/10.1007/s00216-019-01952-6
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author Leivo, Janne
Kivimäki, Laura
Juntunen, Etvi
Pettersson, Kim
Lamminmäki, Urpo
author_facet Leivo, Janne
Kivimäki, Laura
Juntunen, Etvi
Pettersson, Kim
Lamminmäki, Urpo
author_sort Leivo, Janne
collection PubMed
description Detection of circulatory estradiol has widespread use in various clinical applications. Particularly, the use of estradiol-specific antibodies in immunoassays is routinely used, mainly due to the cost efficiency and simplicity of the sample handling process. However, the circulatory levels of estradiol can be extremely low in some conditions, and beyond the current detection limit of existing competitive immunoassays. We describe the generation of anti-immunocomplex specific antibodies derived from synthetic antibody repertoire and the development of high-performance non-competitive immunoassay for the detection of estradiol. Phage display selections were used to isolate new antibodies from synthetic antibody library with the use of existing estradiol specific Fab fragment. The found antibodies were consecutively used to set up a time-resolved fluorescence-based immunoassay (TRFIA), which can be used to detect estradiol with exceptional sensitivity and specificity. The limit of detection and EC50 were shown to be 3.0 pg mL(−1) and 32.4 pg mL(−1) respectively. [Figure: see text]
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spelling pubmed-67042592019-09-06 Development of anti-immunocomplex specific antibodies and non-competitive time-resolved fluorescence immunoassay for the detection of estradiol Leivo, Janne Kivimäki, Laura Juntunen, Etvi Pettersson, Kim Lamminmäki, Urpo Anal Bioanal Chem Communication Detection of circulatory estradiol has widespread use in various clinical applications. Particularly, the use of estradiol-specific antibodies in immunoassays is routinely used, mainly due to the cost efficiency and simplicity of the sample handling process. However, the circulatory levels of estradiol can be extremely low in some conditions, and beyond the current detection limit of existing competitive immunoassays. We describe the generation of anti-immunocomplex specific antibodies derived from synthetic antibody repertoire and the development of high-performance non-competitive immunoassay for the detection of estradiol. Phage display selections were used to isolate new antibodies from synthetic antibody library with the use of existing estradiol specific Fab fragment. The found antibodies were consecutively used to set up a time-resolved fluorescence-based immunoassay (TRFIA), which can be used to detect estradiol with exceptional sensitivity and specificity. The limit of detection and EC50 were shown to be 3.0 pg mL(−1) and 32.4 pg mL(−1) respectively. [Figure: see text] Springer Berlin Heidelberg 2019-06-08 2019 /pmc/articles/PMC6704259/ /pubmed/31177333 http://dx.doi.org/10.1007/s00216-019-01952-6 Text en © The Author(s) 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Communication
Leivo, Janne
Kivimäki, Laura
Juntunen, Etvi
Pettersson, Kim
Lamminmäki, Urpo
Development of anti-immunocomplex specific antibodies and non-competitive time-resolved fluorescence immunoassay for the detection of estradiol
title Development of anti-immunocomplex specific antibodies and non-competitive time-resolved fluorescence immunoassay for the detection of estradiol
title_full Development of anti-immunocomplex specific antibodies and non-competitive time-resolved fluorescence immunoassay for the detection of estradiol
title_fullStr Development of anti-immunocomplex specific antibodies and non-competitive time-resolved fluorescence immunoassay for the detection of estradiol
title_full_unstemmed Development of anti-immunocomplex specific antibodies and non-competitive time-resolved fluorescence immunoassay for the detection of estradiol
title_short Development of anti-immunocomplex specific antibodies and non-competitive time-resolved fluorescence immunoassay for the detection of estradiol
title_sort development of anti-immunocomplex specific antibodies and non-competitive time-resolved fluorescence immunoassay for the detection of estradiol
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6704259/
https://www.ncbi.nlm.nih.gov/pubmed/31177333
http://dx.doi.org/10.1007/s00216-019-01952-6
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