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Flow-Based Chemiluminescence Microarrays as Screening Platform for Affinity Binders to Capture and Elute Bacteria

Affinity describes the non-covalent but selective interaction between an affinity binder (e.g., proteins, antibiotics, or antibodies) and its counterpart (e.g., bacteria). These affinity binders can serve to detect bacteria and respond to the need for selective concentration via affinity chromatogra...

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Detalles Bibliográficos
Autores principales: Neumair, Julia, Elsner, Martin, Seidel, Michael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9693076/
https://www.ncbi.nlm.nih.gov/pubmed/36433201
http://dx.doi.org/10.3390/s22228606
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author Neumair, Julia
Elsner, Martin
Seidel, Michael
author_facet Neumair, Julia
Elsner, Martin
Seidel, Michael
author_sort Neumair, Julia
collection PubMed
description Affinity describes the non-covalent but selective interaction between an affinity binder (e.g., proteins, antibiotics, or antibodies) and its counterpart (e.g., bacteria). These affinity binders can serve to detect bacteria and respond to the need for selective concentration via affinity chromatography for trace analysis. By changing the pH value or salt and protein contents, affinity bindings can be reversed, and bacteria can be recovered for characterisation. Analytical microarrays use multiple affinity binders immobilised on the surface in a distinct pattern, which immensely reduces screening time for the discovery of superior binding motifs. Here, flow-based microarray systems can inform not only about binding, but also about desorption. In this work, we pioneer a screening assay for affinity binders against both gram-positive and negative bacteria based on an automated flow-based chemiluminescence (CL) microarray. Biotinylation of model organisms E. coli and E. faecalis enabled labelling with horseradish-peroxidase-coupled streptavidin, and detection with CL. Polymyxin B, an antibiotic against gram-negative bacteria, was found to bind both E. coli and E. faecalis. Simultaneous screening for desorption methods unexpectedly revealed methyl alpha-D-mannopyranoside as a promising buffer for desorption from Polymyxin B. This proof-of-principle study shows that our new platform greatly facilitates the screening of new affinity binders against bacteria, with promise for future automation.
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spelling pubmed-96930762022-11-26 Flow-Based Chemiluminescence Microarrays as Screening Platform for Affinity Binders to Capture and Elute Bacteria Neumair, Julia Elsner, Martin Seidel, Michael Sensors (Basel) Article Affinity describes the non-covalent but selective interaction between an affinity binder (e.g., proteins, antibiotics, or antibodies) and its counterpart (e.g., bacteria). These affinity binders can serve to detect bacteria and respond to the need for selective concentration via affinity chromatography for trace analysis. By changing the pH value or salt and protein contents, affinity bindings can be reversed, and bacteria can be recovered for characterisation. Analytical microarrays use multiple affinity binders immobilised on the surface in a distinct pattern, which immensely reduces screening time for the discovery of superior binding motifs. Here, flow-based microarray systems can inform not only about binding, but also about desorption. In this work, we pioneer a screening assay for affinity binders against both gram-positive and negative bacteria based on an automated flow-based chemiluminescence (CL) microarray. Biotinylation of model organisms E. coli and E. faecalis enabled labelling with horseradish-peroxidase-coupled streptavidin, and detection with CL. Polymyxin B, an antibiotic against gram-negative bacteria, was found to bind both E. coli and E. faecalis. Simultaneous screening for desorption methods unexpectedly revealed methyl alpha-D-mannopyranoside as a promising buffer for desorption from Polymyxin B. This proof-of-principle study shows that our new platform greatly facilitates the screening of new affinity binders against bacteria, with promise for future automation. MDPI 2022-11-08 /pmc/articles/PMC9693076/ /pubmed/36433201 http://dx.doi.org/10.3390/s22228606 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
Neumair, Julia
Elsner, Martin
Seidel, Michael
Flow-Based Chemiluminescence Microarrays as Screening Platform for Affinity Binders to Capture and Elute Bacteria
title Flow-Based Chemiluminescence Microarrays as Screening Platform for Affinity Binders to Capture and Elute Bacteria
title_full Flow-Based Chemiluminescence Microarrays as Screening Platform for Affinity Binders to Capture and Elute Bacteria
title_fullStr Flow-Based Chemiluminescence Microarrays as Screening Platform for Affinity Binders to Capture and Elute Bacteria
title_full_unstemmed Flow-Based Chemiluminescence Microarrays as Screening Platform for Affinity Binders to Capture and Elute Bacteria
title_short Flow-Based Chemiluminescence Microarrays as Screening Platform for Affinity Binders to Capture and Elute Bacteria
title_sort flow-based chemiluminescence microarrays as screening platform for affinity binders to capture and elute bacteria
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9693076/
https://www.ncbi.nlm.nih.gov/pubmed/36433201
http://dx.doi.org/10.3390/s22228606
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