Rapid profiling of the antigen regions recognized by serum antibodies using massively parallel sequencing of antigen-specific libraries.

Handle URI:
http://hdl.handle.net/10754/596814
Title:
Rapid profiling of the antigen regions recognized by serum antibodies using massively parallel sequencing of antigen-specific libraries.
Authors:
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
Abstract:
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.
Citation:
Domina M, Lanza Cariccio V, Benfatto S, D’Aliberti D, Venza M, et al. (2014) Rapid Profiling of the Antigen Regions Recognized by Serum Antibodies Using Massively Parallel Sequencing of Antigen-Specific Libraries. PLoS ONE 9: e114159. Available: http://dx.doi.org/10.1371/journal.pone.0114159.
Publisher:
Public Library of Science (PLoS)
Journal:
PLoS ONE
KAUST Grant Number:
KUK I1-012-43
Issue Date:
4-Dec-2014
DOI:
10.1371/journal.pone.0114159
PubMed ID:
25473968
PubMed Central ID:
PMC4256389
Type:
Article
ISSN:
1932-6203
Sponsors:
Funding for this work came from CLUSTER MEDINTECH (Project CTN01_00177_962865), PANLab (PON a3_00166), King Abdullah University of Science and Technology (Award KUK I1-012-43), Progetti di Ricerca di Rilevante Interesse Nazionale (20108XYHJS), and the Flagship EPIGEN. All funding for this project is gratefully acknowledged. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Appears in Collections:
Publications Acknowledging KAUST Support

Full metadata record

DC FieldValue Language
dc.contributor.authorDomina, Mariaen
dc.contributor.authorLanza Cariccio, Veronicaen
dc.contributor.authorBenfatto, Salvatoreen
dc.contributor.authorD'Aliberti, Deborahen
dc.contributor.authorVenza, Marioen
dc.contributor.authorBorgogni, Ericaen
dc.contributor.authorCastellino, Floraen
dc.contributor.authorBiondo, Carmeloen
dc.contributor.authorD'Andrea, Danielen
dc.contributor.authorGrassi, Luigien
dc.contributor.authorTramontano, Annaen
dc.contributor.authorTeti, Giuseppeen
dc.contributor.authorFelici, Francoen
dc.contributor.authorBeninati, Concettaen
dc.date.accessioned2016-02-21T08:51:11Zen
dc.date.available2016-02-21T08:51:11Zen
dc.date.issued2014-12-04en
dc.identifier.citationDomina M, Lanza Cariccio V, Benfatto S, D’Aliberti D, Venza M, et al. (2014) Rapid Profiling of the Antigen Regions Recognized by Serum Antibodies Using Massively Parallel Sequencing of Antigen-Specific Libraries. PLoS ONE 9: e114159. Available: http://dx.doi.org/10.1371/journal.pone.0114159.en
dc.identifier.issn1932-6203en
dc.identifier.pmid25473968en
dc.identifier.doi10.1371/journal.pone.0114159en
dc.identifier.urihttp://hdl.handle.net/10754/596814en
dc.description.abstractThere 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.en
dc.description.sponsorshipFunding for this work came from CLUSTER MEDINTECH (Project CTN01_00177_962865), PANLab (PON a3_00166), King Abdullah University of Science and Technology (Award KUK I1-012-43), Progetti di Ricerca di Rilevante Interesse Nazionale (20108XYHJS), and the Flagship EPIGEN. All funding for this project is gratefully acknowledged. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.en
dc.publisherPublic Library of Science (PLoS)en
dc.rightsThis is an open-access article distributed under the terms of the , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en
dc.subject.meshHigh-Throughput Nucleotide Sequencingen
dc.titleRapid profiling of the antigen regions recognized by serum antibodies using massively parallel sequencing of antigen-specific libraries.en
dc.typeArticleen
dc.identifier.journalPLoS ONEen
dc.identifier.pmcidPMC4256389en
dc.contributor.institutionSPGMB Department, University of Messina, Messina, Italy.en
dc.contributor.institutionSSMCSO Department, University of Messina, Messina, Italy.en
dc.contributor.institutionNovartis Vaccines and Diagnostics, Siena, Italy.en
dc.contributor.institutionDepartment of Physics, Sapienza University, Rome, Italy.en
dc.contributor.institutionSPGMB Department, University of Messina, Messina, Italy; Charybdis Vaccines Srl, Messina, Italy.en
dc.contributor.institutionDepartment of Biosciences and Territory, University of Molise, Pesche, Isernia, Italy.en
kaust.grant.numberKUK I1-012-43en

Related articles on PubMed

All Items in KAUST are protected by copyright, with all rights reserved, unless otherwise indicated.