Determination of protein-derived epitopes by mass spectrometry

Hager-Braun, Christine; Tomer, Kenneth B.

doi:10.1586/14789450.2.5.745

Cited by 69 publications

(60 citation statements)

References 72 publications

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“…The rate at which backbone amide hydrogens exchange in solution is directly dependent on the dynamics and structure of the protein. Therefore, regions embedded within the structure or occluded by the presence of a ligand will exchange more slowly than regions fully exposed to the solvent (24).…”

Section: Recombinant C-terminal Domain Of Fhbp Binds Mab 12c1 With Lowermentioning

confidence: 99%

Defining a protective epitope on factor H binding protein, a key meningococcal virulence factor and vaccine antigen

Malito

Faleri

Surdo

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

126

166

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Mapping of epitopes recognized by functional monoclonal antibodies (mAbs) is essential for understanding the nature of immune responses and designing improved vaccines, therapeutics, and diagnostics. In recent years, identification of B-cell epitopes targeted by neutralizing antibodies has facilitated the design of peptide-based vaccines against highly variable pathogens like HIV, respiratory syncytial virus, and Helicobacter pylori; however, none of these products has yet progressed into clinical stages. Linear epitopes identified by conventional mapping techniques only partially reflect the immunogenic properties of the epitope in its natural conformation, thus limiting the success of this approach. To investigate antigen-antibody interactions and assess the potential of the most common epitope mapping techniques, we generated a series of mAbs against factor H binding protein (fHbp), a key virulence factor and vaccine antigen of Neisseria meningitidis. The interaction of fHbp with the bactericidal mAb 12C1 was studied by various epitope mapping methods. Although a 12-residue epitope in the C terminus of fHbp was identified by both Peptide Scanning and Phage Display Library screening, other approaches, such as hydrogen/ deuterium exchange mass spectrometry (MS) and X-ray crystallography, showed that mAb 12C1 occupies an area of ∼1,000 Å 2 on fHbp, including >20 fHbp residues distributed on both N-and C-terminal domains. Collectively, these data show that linear epitope mapping techniques provide useful but incomplete descriptions of B-cell epitopes, indicating that increased efforts to fully characterize antigen-antibody interfaces are required to understand and design effective immunogens.meningococcus | structure | surface plasmon resonance | structural mass spectrometry | antigen-antibody complex

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Section: Recombinant C-terminal Domain Of Fhbp Binds Mab 12c1 With Lowermentioning

confidence: 99%

Defining a protective epitope on factor H binding protein, a key meningococcal virulence factor and vaccine antigen

Malito

Faleri

Surdo

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

126

166

View full text Add to dashboard Cite

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“…4E10 binds to a highly conserved linear epitope on MPER 671 NWF(D/N)IT 676 (161). Crystallographic structures of the epitope have been characterized and form a large helical shape with the important residues on the same side of the helix (44). The antigen reacts only with residues of the base and center of the loop of CDRH3.…”

Section: Gp41 Immunogenicitymentioning

confidence: 99%

“…The 671 NWFDITNWLWYI K 683 sequence is optimal for 4E10 recognition. In resistant strains, the pattern NWF(N/D)IT indicates that the 4E10 epitope is complex and would even be discontinuous (44). Mass spectrometry studies reveal that 4E10 binds to the N terminus of gp120 and gp41 on the native conformation (130).…”

Section: Gp41 Immunogenicitymentioning

confidence: 99%

Role of Human Immunodeficiency Virus Type 1 Envelope Structure in the Induction of Broadly Neutralizing Antibodies

Benjelloun

Lawrence

Verrier

et al. 2012

J Virol

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c Very soon after the discovery of neutralizing antibodies (NAbs) toward human immunodeficiency virus type 1 (HIV-1) infection, it became apparent that characterization of these NAbs would be an important step in finding a cure for or a vaccine to eradicate HIV-1. Since the initial description of broadly cross-clade NAbs naturally produced in HIV-1 patients, numerous studies have described new viral targets for these antibodies. More recently, studies concerning new groups of patients able to control their viremia, such as long-term nonprogressors (LTNPs) or elite controllers, have described the generation of numerous envelope-targeted NAbs. Recent studies have marked a new stage in research on NAbs with the description of antibodies obtained from a worldwide screening of HIV-positive patients. These studies have permitted the discovery of NAb families with great potential for both neutralization and neutralization breadth, such as PG, PGT, CH, and highly active agonistic anti-CD4 binding site antibodies (HAADs), of which VRC01 and its variants are members. These antibodies are able to neutralize more than 80% of circulating strains without any autoreactivity and can be rapidly integrated into clinical trials in order to test their protective potential. In this review, we will focus on new insights into HIV-1 envelope structure and their implications for the generation of potent NAbs.

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“…When the antigen-antibody complex forms, the interface between the binding partners can occlude solvent accessibility, reducing the exchange rate. 16 Epitope mapping by HDX-MS was carried out in 2 parallel steps: deuterium incorporation was performed on NadA alone (reference experiment) and on the antigen-antibody complex, as previously described.…”

Section: Epitope Mapping Using Hdx-msmentioning

confidence: 99%

Phage display revisited: Epitope mapping of a monoclonal antibody directed againstNeisseria meningitidisadhesin A using the PROFILER technology

Domina

Benfatto

et al. 2016

mAbs

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There is a strong need for rapid and reliable epitope mapping methods that can keep pace with the isolation of increasingly larger numbers of mAbs. We describe here the identification of a conformational epitope using Phage-based Representation OF ImmunoLigand Epitope Repertoire (PROFILER), a recently developed high-throughput method based on deep sequencing of antigen-specific lambda phagedisplayed libraries. A novel bactericidal monoclonal antibody (mAb 9F11) raised against Neisseria meningitidis adhesin A (NadA), an important component of the Bexsero Ò anti-meningococcal vaccine, was used to evaluate the technique in comparison with other epitope mapping methods. The PROFILER technology readily identified NadA fragments that were capable of fully recapitulating the reactivity of the entire antigen against mAb 9F11. Further analysis of these fragments using mutagenesis and hydrogendeuterium exchange mass-spectrometry allowed us to identify the binding site of mAb 9F11 (A250-D274) and an adjoining sequence (V275-H312) that was also required for the full functional reconstitution of the epitope. These data suggest that, by virtue of its ability to detect a great variety of immunoreactive antigen fragments in phage-displayed libraries, the PROFILER technology can rapidly and reliably identify epitope-containing regions and provide, in addition, useful clues for the functional characterization of conformational mAb epitopes.

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Determination of protein-derived epitopes by mass spectrometry

Cited by 69 publications

References 72 publications

Defining a protective epitope on factor H binding protein, a key meningococcal virulence factor and vaccine antigen

Defining a protective epitope on factor H binding protein, a key meningococcal virulence factor and vaccine antigen

Role of Human Immunodeficiency Virus Type 1 Envelope Structure in the Induction of Broadly Neutralizing Antibodies

Phage display revisited: Epitope mapping of a monoclonal antibody directed againstNeisseria meningitidisadhesin A using the PROFILER technology

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