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Investigation of Conditions for Capture of Live Legionella pneumophila with Polyclonal and Recombinant Antibodies

Since Legionella pneumophila has caused punctual epidemics through various water systems, the need for a biosensor for fast and accurate detection of pathogenic bacteria in industrial and environmental water has increased. In this report, we evaluated conditions for the capture of live L. pneumophil...

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Autores principales: Paladines, Lucas, Hassen, Walid M., Chawich, Juliana, Dübel, Stefan, Lévesque, Simon, Dubowski, Jan J., Frost, Eric H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9221320/
https://www.ncbi.nlm.nih.gov/pubmed/35735528
http://dx.doi.org/10.3390/bios12060380
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author Paladines, Lucas
Hassen, Walid M.
Chawich, Juliana
Dübel, Stefan
Lévesque, Simon
Dubowski, Jan J.
Frost, Eric H.
author_facet Paladines, Lucas
Hassen, Walid M.
Chawich, Juliana
Dübel, Stefan
Lévesque, Simon
Dubowski, Jan J.
Frost, Eric H.
author_sort Paladines, Lucas
collection PubMed
description Since Legionella pneumophila has caused punctual epidemics through various water systems, the need for a biosensor for fast and accurate detection of pathogenic bacteria in industrial and environmental water has increased. In this report, we evaluated conditions for the capture of live L. pneumophila on a surface by polyclonal antibodies (pAb) and recombinant antibodies (recAb) targeting the bacterial lipopolysaccharide. Using immunoassay and PCR quantification, we demonstrated that, when exposed to live L. pneumophila in PBS or in a mixture containing other non-target bacteria, recAb captured one third fewer L. pneumophila than pAb, but with a 40% lower standard deviation, even when using the same batch of pAb. The presence of other bacteria did not interfere with capture nor increase background by either Ab. Increased reproducibility, as manifested by low standard deviation, is a characteristic that is coveted for biosensing. Hence, the recAb provided a better choice for immune adhesion in biosensors even though it was slightly less sensitive than pAb. Polyclonal or recombinant antibodies can specifically capture large targets such as whole bacteria, and this opens the door to multiple biosensor approaches where any of the components of the bacteria can then be measured for detection or characterisation.
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spelling pubmed-92213202022-06-24 Investigation of Conditions for Capture of Live Legionella pneumophila with Polyclonal and Recombinant Antibodies Paladines, Lucas Hassen, Walid M. Chawich, Juliana Dübel, Stefan Lévesque, Simon Dubowski, Jan J. Frost, Eric H. Biosensors (Basel) Article Since Legionella pneumophila has caused punctual epidemics through various water systems, the need for a biosensor for fast and accurate detection of pathogenic bacteria in industrial and environmental water has increased. In this report, we evaluated conditions for the capture of live L. pneumophila on a surface by polyclonal antibodies (pAb) and recombinant antibodies (recAb) targeting the bacterial lipopolysaccharide. Using immunoassay and PCR quantification, we demonstrated that, when exposed to live L. pneumophila in PBS or in a mixture containing other non-target bacteria, recAb captured one third fewer L. pneumophila than pAb, but with a 40% lower standard deviation, even when using the same batch of pAb. The presence of other bacteria did not interfere with capture nor increase background by either Ab. Increased reproducibility, as manifested by low standard deviation, is a characteristic that is coveted for biosensing. Hence, the recAb provided a better choice for immune adhesion in biosensors even though it was slightly less sensitive than pAb. Polyclonal or recombinant antibodies can specifically capture large targets such as whole bacteria, and this opens the door to multiple biosensor approaches where any of the components of the bacteria can then be measured for detection or characterisation. MDPI 2022-05-31 /pmc/articles/PMC9221320/ /pubmed/35735528 http://dx.doi.org/10.3390/bios12060380 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Paladines, Lucas
Hassen, Walid M.
Chawich, Juliana
Dübel, Stefan
Lévesque, Simon
Dubowski, Jan J.
Frost, Eric H.
Investigation of Conditions for Capture of Live Legionella pneumophila with Polyclonal and Recombinant Antibodies
title Investigation of Conditions for Capture of Live Legionella pneumophila with Polyclonal and Recombinant Antibodies
title_full Investigation of Conditions for Capture of Live Legionella pneumophila with Polyclonal and Recombinant Antibodies
title_fullStr Investigation of Conditions for Capture of Live Legionella pneumophila with Polyclonal and Recombinant Antibodies
title_full_unstemmed Investigation of Conditions for Capture of Live Legionella pneumophila with Polyclonal and Recombinant Antibodies
title_short Investigation of Conditions for Capture of Live Legionella pneumophila with Polyclonal and Recombinant Antibodies
title_sort investigation of conditions for capture of live legionella pneumophila with polyclonal and recombinant antibodies
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9221320/
https://www.ncbi.nlm.nih.gov/pubmed/35735528
http://dx.doi.org/10.3390/bios12060380
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