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Rapid Profiling of the Antigen Regions Recognized by Serum Antibodies Using Massively Parallel Sequencing of Antigen-Specific Libraries
There is a need for techniques capable of identifying the antigenic epitopes targeted by polyclonal antibody responses during deliberate or natural immunization. Although successful, traditional phage library screening is laborious and can map only some of the epitopes. To accelerate and improve epi...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4256389/ https://www.ncbi.nlm.nih.gov/pubmed/25473968 http://dx.doi.org/10.1371/journal.pone.0114159 |
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author | Domina, Maria Lanza Cariccio, Veronica Benfatto, Salvatore D'Aliberti, Deborah Venza, Mario Borgogni, Erica Castellino, Flora Biondo, Carmelo D'Andrea, Daniel Grassi, Luigi Tramontano, Anna Teti, Giuseppe Felici, Franco Beninati, Concetta |
author_facet | Domina, Maria Lanza Cariccio, Veronica Benfatto, Salvatore D'Aliberti, Deborah Venza, Mario Borgogni, Erica Castellino, Flora Biondo, Carmelo D'Andrea, Daniel Grassi, Luigi Tramontano, Anna Teti, Giuseppe Felici, Franco Beninati, Concetta |
author_sort | Domina, Maria |
collection | PubMed |
description | There is a need for techniques capable of identifying the antigenic epitopes targeted by polyclonal antibody responses during deliberate or natural immunization. Although successful, traditional phage library screening is laborious and can map only some of the epitopes. To accelerate and improve epitope identification, we have employed massive sequencing of phage-displayed antigen-specific libraries using the Illumina MiSeq platform. This enabled us to precisely identify the regions of a model antigen, the meningococcal NadA virulence factor, targeted by serum antibodies in vaccinated individuals and to rank hundreds of antigenic fragments according to their immunoreactivity. We found that next generation sequencing can significantly empower the analysis of antigen-specific libraries by allowing simultaneous processing of dozens of library/serum combinations in less than two days, including the time required for antibody-mediated library selection. Moreover, compared with traditional plaque picking, the new technology (named Phage-based Representation OF Immuno-Ligand Epitope Repertoire or PROFILER) provides superior resolution in epitope identification. PROFILER seems ideally suited to streamline and guide rational antigen design, adjuvant selection, and quality control of newly produced vaccines. Furthermore, this method is also susceptible to find important applications in other fields covered by traditional quantitative serology. |
format | Online Article Text |
id | pubmed-4256389 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-42563892014-12-11 Rapid Profiling of the Antigen Regions Recognized by Serum Antibodies Using Massively Parallel Sequencing of Antigen-Specific Libraries Domina, Maria Lanza Cariccio, Veronica Benfatto, Salvatore D'Aliberti, Deborah Venza, Mario Borgogni, Erica Castellino, Flora Biondo, Carmelo D'Andrea, Daniel Grassi, Luigi Tramontano, Anna Teti, Giuseppe Felici, Franco Beninati, Concetta PLoS One Research Article There is a need for techniques capable of identifying the antigenic epitopes targeted by polyclonal antibody responses during deliberate or natural immunization. Although successful, traditional phage library screening is laborious and can map only some of the epitopes. To accelerate and improve epitope identification, we have employed massive sequencing of phage-displayed antigen-specific libraries using the Illumina MiSeq platform. This enabled us to precisely identify the regions of a model antigen, the meningococcal NadA virulence factor, targeted by serum antibodies in vaccinated individuals and to rank hundreds of antigenic fragments according to their immunoreactivity. We found that next generation sequencing can significantly empower the analysis of antigen-specific libraries by allowing simultaneous processing of dozens of library/serum combinations in less than two days, including the time required for antibody-mediated library selection. Moreover, compared with traditional plaque picking, the new technology (named Phage-based Representation OF Immuno-Ligand Epitope Repertoire or PROFILER) provides superior resolution in epitope identification. PROFILER seems ideally suited to streamline and guide rational antigen design, adjuvant selection, and quality control of newly produced vaccines. Furthermore, this method is also susceptible to find important applications in other fields covered by traditional quantitative serology. Public Library of Science 2014-12-04 /pmc/articles/PMC4256389/ /pubmed/25473968 http://dx.doi.org/10.1371/journal.pone.0114159 Text en © 2014 Domina et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Domina, Maria Lanza Cariccio, Veronica Benfatto, Salvatore D'Aliberti, Deborah Venza, Mario Borgogni, Erica Castellino, Flora Biondo, Carmelo D'Andrea, Daniel Grassi, Luigi Tramontano, Anna Teti, Giuseppe Felici, Franco Beninati, Concetta Rapid Profiling of the Antigen Regions Recognized by Serum Antibodies Using Massively Parallel Sequencing of Antigen-Specific Libraries |
title | Rapid Profiling of the Antigen Regions Recognized by Serum Antibodies Using Massively Parallel Sequencing of Antigen-Specific Libraries |
title_full | Rapid Profiling of the Antigen Regions Recognized by Serum Antibodies Using Massively Parallel Sequencing of Antigen-Specific Libraries |
title_fullStr | Rapid Profiling of the Antigen Regions Recognized by Serum Antibodies Using Massively Parallel Sequencing of Antigen-Specific Libraries |
title_full_unstemmed | Rapid Profiling of the Antigen Regions Recognized by Serum Antibodies Using Massively Parallel Sequencing of Antigen-Specific Libraries |
title_short | Rapid Profiling of the Antigen Regions Recognized by Serum Antibodies Using Massively Parallel Sequencing of Antigen-Specific Libraries |
title_sort | rapid profiling of the antigen regions recognized by serum antibodies using massively parallel sequencing of antigen-specific libraries |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4256389/ https://www.ncbi.nlm.nih.gov/pubmed/25473968 http://dx.doi.org/10.1371/journal.pone.0114159 |
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