Mapping the Epitopes of Antibodies

Ladner, Robert C.

doi:10.1080/02648725.2007.10648092

Cited by 45 publications

(34 citation statements)

References 76 publications

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“…Such information can be derived from several epitope mapping approaches (28). However, most conventional techniques are limited, because they detect interactions between a mAb and a single short continuous stretch of residues of the antigen, which may or may not adopt relevant secondary or tertiary structure, thus being unable to faithfully mimic complex protein conformations.…”

Section: Discussionmentioning

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: Discussionmentioning

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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“…As predicted from a competitive ELISA, three epitopes of sfGFP are targeted by these seven nanobodies; I(01, 02), II(03, 06) and III(04, 07, 08), and the epitopes I and II seems to be so close on the surface of sfGFP that both were targeted by a mouse anti-GFP antibody. Epitope mapping is an important step in characterizing antibodies that bind a protein target [37].…”

Section: Discussionmentioning

confidence: 99%

Characterization of camel nanobodies specific for superfolder GFP fusion proteins

et al. 2014

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Superfolder green fluorescent protein (sfGFP) is a fusion tag which plays a dual role in monitoring and purifying the recombinant fusion proteins using specific binders. Nanobodies are the smallest intact antigen binding fragments derived from heavy chain-only antibodies (HCAbs) occurring in camelids. They are produced as recombinant proteins in E. coli and have different biotechnological applications, including the detection and purification of their specific antigens. To produce anti-sfGFP specific nanobodies, an adult one-humped camel was successfully immunized and immune response was evaluated by ELISA, which showed an active participation of HCAbs in this response. A relatively large nanobody "immune" library of 5 × 10(8) individual transformants, with 87.5 % positivity, was prepared from the blood of the immunized camel. Phage display biopanning on this nanobody library resulted in the isolation of seven anti-sfGFP specific nanobodies, referred to as NbsfGFP01, 02, 03, 04, 06, 07 and 08. These nanobodies were able to recognize sfGFP tag as free or in fusion with growth hormone in ELISA and immuno-blotting. Furthermore, they showed important apparent affinities in the detection and capture of sfGFP by ELISA, and they targeted three different epitopes on the surface of their antigen. The interesting characteristics of these molecular binders make them valuable tools for more in-depth structural and functional studies related to sfGFP fusion proteins.

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“…Helicobacter pylori could trigger infections in the epithelial layer of human gastrointestinal tract, particularly in the gastric antrum. This bacterium is rarely isolated from the rectum, esophagus, bowel, or blood (15). Urease is considered a significant pathogenic indicator of H. pylori.…”

Section: Discussionmentioning

confidence: 99%