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Epitope mapping of a monoclonal antibody directed against Neisserial Heparin Binding Antigen

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Mus musculus Neisseria meningitidis serogroup B Neisseria meningitidis

PROVIDER: GSE81379 | GEO | 2016/05/27

SECONDARY ACCESSION(S): PRJNA321411

REPOSITORIES: GEO

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Epitope mapping of a monoclonal antibody directed against Neisserial Heparin Binding Antigen by peptide microarray

Project description:The aim of the study was to determine the epitope targeted by 31E10/E7 mouse monoclonal antibody (mAb) and the cross-reactivity to linear peptide epitopes of 10 different Neisserial Heparin Binding Antigen (NHBA) variants. mAb 31E10/E7 was diluted at 1:200 and incubated on a custom PepStar Peptide Microarray platform printed with 560 different peptides in three independent experiments.

2016-05-27 | GSE81377 | GEO

Epitope mapping of a monoclonal antibody directed against Neisserial Heparin Binding Antigen by protein microarray

Project description:The aim of the study was to determine the epitope targeted by 31E10/E7 mouse monoclonal antibody (mAb). mAb 31E10/E7 was diluted at 1:2000 and incubated on a non-commercial Protein Microarray platform printed with NHBA specific recombinant protein fragments and full length NHBA of different variants.

2016-05-27 | GSE81378 | GEO

Epitope mapping of 4 monoclonal antibodies directed against Neisserial Heparin Binding Antigen by peptide microarray

Project description:The aim of the study was to determine the epitope targeted by four different HumAbs and the cross-reactivity to linear peptide epitopes of 10 different Neisserial Heparin Binding Antigen (NHBA) variants. the HumAbs were diluted at 1:60 and incubated on a custom PepStar Peptide Microarray platform printed with 561 different peptides.

2018-03-07 | GSE98882 | GEO

Epitope Mapping of a Monoclonal Antibody Directed against Neisserial Heparin Binding Antigen Using Next Generation Sequencing of Antigen-Specific Libraries.

Project 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.

| S-EPMC4980009 | biostudies-literature

Epitope mapping of a monoclonal antibody directed against Neisserial Heparin Binding Antigen

Project description:Epitope mapping of a monoclonal antibody directed against Neisserial Heparin Binding Antigen

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Epitope mapping of 11 monoclonal antibodies directed against Neisserial adesin A by peptide microarray

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2018-03-07 | GSE98881 | GEO

Epitope mapping of 5 monoclonal antibodies directed against meningococcal factor H binding protein by peptide microarray

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2018-03-07 | GSE98879 | GEO

Mapping of the epitope/paratope interactions of a monoclonal antibody directed against adenosine 3',5'-monophosphate.

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Epitope mapping of human antibodies directed against 4CmenB protein components

Project description:This SuperSeries is composed of the SubSeries listed below.

2018-03-07 | GSE98883 | GEO

Epitope-directed monoclonal antibody production using a mixed antigen cocktail facilitates antibody characterization and validation.

Project description:High quality, well-validated antibodies are needed to mitigate irreproducibility and clarify conflicting data in science. We describe an epitope-directed monoclonal antibody (mAb) production method that addresses issues of antibody quality, validation and utility. The workflow is illustrated by generating mAbs against multiple in silico-predicted epitopes on human ankyrin repeat domain 1 (hANKRD1) in a single hybridoma production cycle. Antigenic peptides (13-24 residues long) presented as three-copy inserts on the surface exposed loop of a thioredoxin carrier produced high affinity mAbs that are reactive to native and denatured hANKRD1. ELISA assay miniaturization afforded by novel DEXT microplates allowed rapid hybridoma screening with concomitant epitope identification. Antibodies against spatially distant sites on hANKRD1 facilitated validation schemes applicable to two-site ELISA, western blotting and immunocytochemistry. The use of short antigenic peptides of known sequence facilitated direct epitope mapping crucial for antibody characterization. This robust method motivates its ready adoption for other protein targets.

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