Epitope mapping of a monoclonal antibody directed against neisserial heparin binding antigen using next generation sequencing of antigen-specific libraries

Domina, Maria; Cariccio, Veronica Lanza; Benfatto, Salvatore; Venza, Mario; Venza, Isabella; Donnarumma, Danilo; Bartolini, Erika; Borgogni, Erica; Bruttini, Marco; Santini, Laura; Midiri, Angelina; Galbo, Roberta; Romeo, Letizia; Patane', Francesco; Biondo, Carmelo; Norais, Nathalie; Masignani, Vega; Teti, Giuseppe; Felici, Franco; Beninati, Concetta

doi:10.1371/journal.pone.0160702

We explore here the potential of a newly described technology, which is named PROFILER and is based on next generation sequencing of gene-specific lambda phage-displayed libraries, to rapidly and accurately map monoclonal antibody (mAb) epitopes. For this purpose, we used a novel mAb (designated 31E10/E7) directed against Neisserial Heparin-Binding Antigen (NHBA), a component of the anti-group B meningococcus Bexsero1 vaccine. An NHBA phage-displayed library was affinity-selected with mAb 31E10/E7, followed by massive sequencing of the inserts present in antibody-selected phage pools. Insert analysis identified an amino acid stretch (D91-A128) in the N-terminal domain, which was shared by all of the mAb-enriched fragments. Moreover, a recombinant fragment encompassing this sequence could recapitulate the immunoreactivity of the entire NHBA molecule against mAb 31E10/E7. These results were confirmed using a panel of overlapping recombinant fragments derived from the NHBA vaccine variant and a set of chemically synthetized peptides covering the 10 most frequent antigenic variants. Furthermore, hydrogen-deuterium exchange mass-spectrometry analysis of the NHBA-mAb 31E10/E7 complex was also compatible with mapping of the epitope to the D91-A128 region. Collectively, these results indicate that the PROFILER technology can reliably identify epitope-containing antigenic fragments and requires considerably less work, time and reagents than other epitope mapping methods.

Titolo:	Epitope mapping of a monoclonal antibody directed against neisserial heparin binding antigen using next generation sequencing of antigen-specific libraries
Autori:	DOMINA, MARIA (Primo) Cariccio, Veronica Lanza Benfatto, Salvatore VENZA, Mario VENZA, Isabella Donnarumma, Danilo Bartolini, Erika Borgogni, Erica Bruttini, Marco Santini, Laura MIDIRI, Angelina GALBO, Roberta ROMEO, LETIZIA PATANE', FRANCESCO BIONDO, Carmelo Norais, Nathalie Masignani, Vega TETI, Giuseppe (Corresponding) Felici, Franco BENINATI, Concetta (Ultimo) mostra contributor esterni nascondi contributor esterni
Data di pubblicazione:	2016
Rivista:	PLOS ONE
Abstract:	We explore here the potential of a newly described technology, which is named PROFILER and is based on next generation sequencing of gene-specific lambda phage-displayed libraries, to rapidly and accurately map monoclonal antibody (mAb) epitopes. For this purpose, we used a novel mAb (designated 31E10/E7) directed against Neisserial Heparin-Binding Antigen (NHBA), a component of the anti-group B meningococcus Bexsero1 vaccine. An NHBA phage-displayed library was affinity-selected with mAb 31E10/E7, followed by massive sequencing of the inserts present in antibody-selected phage pools. Insert analysis identified an amino acid stretch (D91-A128) in the N-terminal domain, which was shared by all of the mAb-enriched fragments. Moreover, a recombinant fragment encompassing this sequence could recapitulate the immunoreactivity of the entire NHBA molecule against mAb 31E10/E7. These results were confirmed using a panel of overlapping recombinant fragments derived from the NHBA vaccine variant and a set of chemically synthetized peptides covering the 10 most frequent antigenic variants. Furthermore, hydrogen-deuterium exchange mass-spectrometry analysis of the NHBA-mAb 31E10/E7 complex was also compatible with mapping of the epitope to the D91-A128 region. Collectively, these results indicate that the PROFILER technology can reliably identify epitope-containing antigenic fragments and requires considerably less work, time and reagents than other epitope mapping methods.
Handle:	http://hdl.handle.net/11570/3094558
Appare nelle tipologie:	14.a.1 Articolo su rivista

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