Epitope Mapping of a Monoclonal Antibody Directed against Neisserial Heparin Binding Antigen Using Next Generation Sequencing of Antigen-Specific Libraries.
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 31E...
| Main Authors: | , , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2016-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4980009?pdf=render |
| _version_ | 1828888704082509824 |
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| author | Maria Domina Veronica Lanza Cariccio Salvatore Benfatto Mario Venza Isabella Venza Danilo Donnarumma Erika Bartolini Erica Borgogni Marco Bruttini Laura Santini Angelina Midiri Roberta Galbo Letizia Romeo Francesco Patanè Carmelo Biondo Nathalie Norais Vega Masignani Giuseppe Teti Franco Felici Concetta Beninati |
| author_facet | Maria Domina Veronica Lanza Cariccio Salvatore Benfatto Mario Venza Isabella Venza Danilo Donnarumma Erika Bartolini Erica Borgogni Marco Bruttini Laura Santini Angelina Midiri Roberta Galbo Letizia Romeo Francesco Patanè Carmelo Biondo Nathalie Norais Vega Masignani Giuseppe Teti Franco Felici Concetta Beninati |
| author_sort | Maria Domina |
| collection | DOAJ |
| description | 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 Bexsero® 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. |
| first_indexed | 2024-12-13T12:27:22Z |
| format | Article |
| id | doaj.art-454e679ddbdb49edad4836cb956bfb3e |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-13T12:27:22Z |
| publishDate | 2016-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-454e679ddbdb49edad4836cb956bfb3e2022-12-21T23:46:12ZengPublic Library of Science (PLoS)PLoS ONE1932-62032016-01-01118e016070210.1371/journal.pone.0160702Epitope Mapping of a Monoclonal Antibody Directed against Neisserial Heparin Binding Antigen Using Next Generation Sequencing of Antigen-Specific Libraries.Maria DominaVeronica Lanza CariccioSalvatore BenfattoMario VenzaIsabella VenzaDanilo DonnarummaErika BartoliniErica BorgogniMarco BruttiniLaura SantiniAngelina MidiriRoberta GalboLetizia RomeoFrancesco PatanèCarmelo BiondoNathalie NoraisVega MasignaniGiuseppe TetiFranco FeliciConcetta BeninatiWe 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 Bexsero® 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.http://europepmc.org/articles/PMC4980009?pdf=render |
| spellingShingle | Maria Domina Veronica Lanza Cariccio Salvatore Benfatto Mario Venza Isabella Venza Danilo Donnarumma Erika Bartolini Erica Borgogni Marco Bruttini Laura Santini Angelina Midiri Roberta Galbo Letizia Romeo Francesco Patanè Carmelo Biondo Nathalie Norais Vega Masignani Giuseppe Teti Franco Felici Concetta Beninati Epitope Mapping of a Monoclonal Antibody Directed against Neisserial Heparin Binding Antigen Using Next Generation Sequencing of Antigen-Specific Libraries. PLoS ONE |
| title | Epitope Mapping of a Monoclonal Antibody Directed against Neisserial Heparin Binding Antigen Using Next Generation Sequencing of Antigen-Specific Libraries. |
| title_full | Epitope Mapping of a Monoclonal Antibody Directed against Neisserial Heparin Binding Antigen Using Next Generation Sequencing of Antigen-Specific Libraries. |
| title_fullStr | Epitope Mapping of a Monoclonal Antibody Directed against Neisserial Heparin Binding Antigen Using Next Generation Sequencing of Antigen-Specific Libraries. |
| title_full_unstemmed | Epitope Mapping of a Monoclonal Antibody Directed against Neisserial Heparin Binding Antigen Using Next Generation Sequencing of Antigen-Specific Libraries. |
| title_short | Epitope Mapping of a Monoclonal Antibody Directed against Neisserial Heparin Binding Antigen Using Next Generation Sequencing of Antigen-Specific Libraries. |
| title_sort | epitope mapping of a monoclonal antibody directed against neisserial heparin binding antigen using next generation sequencing of antigen specific libraries |
| url | http://europepmc.org/articles/PMC4980009?pdf=render |
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