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Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies: Application to Antigen 85, a Tuberculosis Biomarker

BACKGROUND: Current diagnostic methods for tuberculosis (TB), a major global health challenge that kills nearly two million people annually, are time-consuming and inadequate. During infection a number of bacterial molecules that play a role in the infective process are released and have been propos...

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Autores principales: Ferrara, Fortunato, Naranjo, Leslie A., Kumar, Sandeep, Gaiotto, Tiziano, Mukundan, Harshini, Swanson, Basil, Bradbury, Andrew R. M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3498134/
https://www.ncbi.nlm.nih.gov/pubmed/23166701
http://dx.doi.org/10.1371/journal.pone.0049535
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author Ferrara, Fortunato
Naranjo, Leslie A.
Kumar, Sandeep
Gaiotto, Tiziano
Mukundan, Harshini
Swanson, Basil
Bradbury, Andrew R. M.
author_facet Ferrara, Fortunato
Naranjo, Leslie A.
Kumar, Sandeep
Gaiotto, Tiziano
Mukundan, Harshini
Swanson, Basil
Bradbury, Andrew R. M.
author_sort Ferrara, Fortunato
collection PubMed
description BACKGROUND: Current diagnostic methods for tuberculosis (TB), a major global health challenge that kills nearly two million people annually, are time-consuming and inadequate. During infection a number of bacterial molecules that play a role in the infective process are released and have been proposed as biomarkers for early TB diagnosis. Antigen 85 (Ag85) is the most abundant secreted TB protein, and a potential target for this diagnostic approach. One of the bottlenecks in the direct detection of such bacterial targets is the availability of robust, sensitive, specific antibodies. METHODS: Using Ag85 as a model, we describe a method to select antibodies against any potential target using a novel combination of phage and yeast display that exploits the advantage of each approach. RESULTS: The efficiency of this approach was attested to by the 111 specific antibodies identified in initial screens. These were assessed for binding to the different Ag85 subunits, affinity, and activity in sandwich assays. CONCLUSIONS: The novelty of this approach lies in the possibility of screening the entire output of a phage antibody selection in a single experiment by yeast display. This can be considered analogous to carrying out a million ELISAs. The monoclonal antibodies (mAbs) identified in this way show high binding affinity and selectivity for the antigens and offer an advantage over traditional mAbs produced by relatively expensive and time consuming techniques. This approach has wide applicability, and the affinity of selected antibodies can be significantly improved, if required.
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spelling pubmed-34981342012-11-19 Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies: Application to Antigen 85, a Tuberculosis Biomarker Ferrara, Fortunato Naranjo, Leslie A. Kumar, Sandeep Gaiotto, Tiziano Mukundan, Harshini Swanson, Basil Bradbury, Andrew R. M. PLoS One Research Article BACKGROUND: Current diagnostic methods for tuberculosis (TB), a major global health challenge that kills nearly two million people annually, are time-consuming and inadequate. During infection a number of bacterial molecules that play a role in the infective process are released and have been proposed as biomarkers for early TB diagnosis. Antigen 85 (Ag85) is the most abundant secreted TB protein, and a potential target for this diagnostic approach. One of the bottlenecks in the direct detection of such bacterial targets is the availability of robust, sensitive, specific antibodies. METHODS: Using Ag85 as a model, we describe a method to select antibodies against any potential target using a novel combination of phage and yeast display that exploits the advantage of each approach. RESULTS: The efficiency of this approach was attested to by the 111 specific antibodies identified in initial screens. These were assessed for binding to the different Ag85 subunits, affinity, and activity in sandwich assays. CONCLUSIONS: The novelty of this approach lies in the possibility of screening the entire output of a phage antibody selection in a single experiment by yeast display. This can be considered analogous to carrying out a million ELISAs. The monoclonal antibodies (mAbs) identified in this way show high binding affinity and selectivity for the antigens and offer an advantage over traditional mAbs produced by relatively expensive and time consuming techniques. This approach has wide applicability, and the affinity of selected antibodies can be significantly improved, if required. Public Library of Science 2012-11-14 /pmc/articles/PMC3498134/ /pubmed/23166701 http://dx.doi.org/10.1371/journal.pone.0049535 Text en https://creativecommons.org/publicdomain/zero/1.0/ This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.
spellingShingle Research Article
Ferrara, Fortunato
Naranjo, Leslie A.
Kumar, Sandeep
Gaiotto, Tiziano
Mukundan, Harshini
Swanson, Basil
Bradbury, Andrew R. M.
Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies: Application to Antigen 85, a Tuberculosis Biomarker
title Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies: Application to Antigen 85, a Tuberculosis Biomarker
title_full Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies: Application to Antigen 85, a Tuberculosis Biomarker
title_fullStr Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies: Application to Antigen 85, a Tuberculosis Biomarker
title_full_unstemmed Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies: Application to Antigen 85, a Tuberculosis Biomarker
title_short Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies: Application to Antigen 85, a Tuberculosis Biomarker
title_sort using phage and yeast display to select hundreds of monoclonal antibodies: application to antigen 85, a tuberculosis biomarker
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3498134/
https://www.ncbi.nlm.nih.gov/pubmed/23166701
http://dx.doi.org/10.1371/journal.pone.0049535
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