Rapid profiling of the antigen regions recognized by serum antibodies using massively parallel sequencing of antigen-specific libraries.
Lanza Cariccio, Veronica
KAUST Grant NumberKUK I1-012-43
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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.
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.
SponsorsFunding 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.
PublisherPublic Library of Science (PLoS)
PubMed Central IDPMC4256389
CollectionsPublications Acknowledging KAUST Support
- Phage display revisited: Epitope mapping of a monoclonal antibody directed against Neisseria meningitidis adhesin A using the PROFILER technology.
- Authors: Cariccio VL, Domina M, Benfatto S, Venza M, Venza I, Faleri A, Bruttini M, Bartolini E, Giuliani MM, Santini L, Brunelli B, Norais N, Borgogni E, Midiri A, Galbo R, Romeo L, Biondo C, Masignani V, Teti G, Felici F, Beninati C
- Issue date: 2016 May-Jun
- Defining a protective epitope on factor H binding protein, a key meningococcal virulence factor and vaccine antigen.
- Authors: Malito E, Faleri A, Lo Surdo P, Veggi D, Maruggi G, Grassi E, Cartocci E, Bertoldi I, Genovese A, Santini L, Romagnoli G, Borgogni E, Brier S, Lo Passo C, Domina M, Castellino F, Felici F, van der Veen S, Johnson S, Lea SM, Tang CM, Pizza M, Savino S, Norais N, Rappuoli R, Bottomley MJ, Masignani V
- Issue date: 2013 Feb 26
- Epitope Mapping of a Monoclonal Antibody Directed against Neisserial Heparin Binding Antigen Using Next Generation Sequencing of Antigen-Specific Libraries.
- Authors: Domina M, Lanza Cariccio V, Benfatto S, Venza M, Venza I, Donnarumma D, Bartolini E, Borgogni E, Bruttini M, Santini L, Midiri A, Galbo R, Romeo L, Patanè F, Biondo C, Norais N, Masignani V, Teti G, Felici F, Beninati C
- Issue date: 2016
- Functional characterization of a monoclonal antibody epitope using a lambda phage display-deep sequencing platform.
- Authors: Domina M, Lanza Cariccio V, Benfatto S, Venza M, Venza I, Borgogni E, Castellino F, Midiri A, Galbo R, Romeo L, Biondo C, Masignani V, Teti G, Felici F, Beninati C
- Issue date: 2016 Aug 17
- Precise and efficient antibody epitope determination through library design, yeast display and next-generation sequencing.
- Authors: Van Blarcom T, Rossi A, Foletti D, Sundar P, Pitts S, Bee C, Melton Witt J, Melton Z, Hasa-Moreno A, Shaughnessy L, Telman D, Zhao L, Cheung WL, Berka J, Zhai W, Strop P, Chaparro-Riggers J, Shelton DL, Pons J, Rajpal A
- Issue date: 2015 Mar 27