Bactericidal Antibody Responses Induced by Meningococcal Recombinant Chimeric Factor H-Binding Protein Vaccines

Beernink, Peter T.; Granoff, Dan M.

doi:10.1128/iai.00033-08

Cited by 64 publications

(82 citation statements)

References 43 publications

(48 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As shown by SPR and FACS analysis, mAb 12C1 was highly specific for fHbp var1. Given the extensive sequence diversity of fHbp, identification of anti-fHbp mAbs displaying variant-restricted specificity is not unusual, particularly for FACS-positive mAbs, because most of the fHbp variability is associated with surface-exposed regions (17). Consequently, only serogroup B meningococcal strains expressing var1 fHbp were sensitive to rabbit complement-mediated bactericidal activity exerted by mAb 12C1.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

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

170

View full text Add to dashboard Cite

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

show abstract

Section: Discussionmentioning

confidence: 99%

“…Several previous epitope mapping studies have been performed on fHbp using NMR spectroscopy (14,15), scanning of synthetic peptide arrays (16), and phage-or yeast-display techniques (17)(18)(19). The interpretation of these studies was facilitated by the 3D structures of fHbp alone (20,21) or in complex with hfH (22).…”

mentioning

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

170

View full text Add to dashboard Cite

show abstract

“…Binding of human FH and anti-FHbp MAbs to purified FHbp was performed as described (9). Briefly the wells of a microtiter plate were coated with 2 μg/mL of purified recombinant FHbp in PBS.…”

Section: Methodsmentioning

confidence: 99%

“…The second licensed vaccine contains FHbp in variant group 1 and three other antigens capable of eliciting protective antibodies (7,8). Other strategies to increase the breadth of protection were to engineer FHbp antigens containing epitopes from different sequence variants (9)(10)(11).…”

mentioning

confidence: 99%

A meningococcal vaccine antigen engineered to increase thermal stability and stabilize protective epitopes

Konar

Pajón

Beernink

2015

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

Self Cite

View full text Add to dashboard Cite

Factor H binding protein (FHbp) is part of two vaccines recently licensed for prevention of sepsis and meningitis caused by serogroup B meningococci. FHbp is classified in three phylogenic variant groups that have limited antigenic cross-reactivity, and FHbp variants in one of the groups have low thermal stability. In the present study, we replaced two amino acid residues, R130 and D133, in a stable FHbp variant with their counterparts (L and G) from a less stable variant. The single and double mutants decreased thermal stability of the amino- (N-) terminal domain compared with the wild-type protein as measured by scanning calorimetry. We introduced the converse substitutions, L130R and G133D, in a less stable wild-type FHbp variant, which increased the transition midpoint (Tm) for the N-terminal domain by 8 and 12 °C; together the substitutions increased the Tm by 21 °C. We determined the crystal structure of the double mutant FHbp to 1.6 Å resolution, which showed that R130 and D133 mediated multiple electrostatic interactions. Monoclonal antibodies specific for FHbp epitopes in the N-terminal domain had higher binding affinity for the recombinant double mutant by surface plasmon resonance and to the mutant expressed on meningococci by flow cytometry. The double mutant also had decreased binding of human complement Factor H, which in previous studies increased the protective antibody responses. The stabilized mutant FHbp thus has the potential to stabilize protective epitopes and increase the protective antibody responses to recombinant FHbp vaccines or native outer membrane vesicle vaccines with overexpressed FHbp.

show abstract

“…Meningococcal strains are classified into 13 serogroups based on the chemical composition of the polysaccharide capsule [2]. Serogroups A, B, C, Y, and W-135 are associated with disease worldwide [3]. Antibiotic therapy is often ineffective to treat meningococcal infections and vaccination is considered an attractive approach for prophylactic purposes [4].…”

Section: Introductionmentioning

confidence: 99%

Cloning, expression and purification of outer membrane protein PorA of Neisseria meningitidis serogroup B

Haghi

Peerayeh

Siadat

et al. 2011

J Infect Dev Ctries

View full text Add to dashboard Cite

Introduction: Neisseria meningitidis is a major causative agent of bacterial septicemia and meningitis in humans. Currently, there are no vaccines to prevent disease caused by strains of N. meningitidis serogroup B. PorA is a major component of the outer membrane of N. meningitidis and functions as a cationic porin. This study aimed to clone and determine the expression of PorA. Methodology: A 1200 bp fragment of porA gene was amplified by PCR from serogroup B N. meningitidis and then cloned into prokaryotic expression vector pET-32a. For expression of recombinant protein, pET32a-porA plasmid was transformed into competent Origami B (DE3) cells. Recombinant protein was overexpressed with isopropythio-beta-D-galctoside (IPTG) and affinity purified by Ni-NTA agarose. SDS-PAGE and western blotting were performed for protein determination and verification. Results: Cloning of porA was confirmed by colony-PCR and enzymatic digestion. In comparison with the corresponding sequences of original genes, the nucleotide sequence homology of the cloned porA gene was 97%. IPTG with a dosage of 1.0 mmol/L could efficiently induce protein expression. SDS-PAGE analysis showed that our constructed prokaryotic expression system pET32a-PorA-Origami efficiently produces a target recombinant protein with a molecular weight of 65 kDa. The recombinant PorA was overexpressed as inclusion bodies and reacted with the serum from a rabbit previously immunized with native outer membrane vesicle. Conclusion: This prokaryotic expression system provides an easy method for producing recombinant PorA and may also be useful for the production of other bacterial outer membrane proteins for vaccine studies.

show abstract

Bactericidal Antibody Responses Induced by Meningococcal Recombinant Chimeric Factor H-Binding Protein Vaccines

Cited by 64 publications

References 43 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

A meningococcal vaccine antigen engineered to increase thermal stability and stabilize protective epitopes

Cloning, expression and purification of outer membrane protein PorA of Neisseria meningitidis serogroup B

Contact Info

Product

Resources

About