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


Abstract

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.

Submission Details

ID: dYNhykwR4

Submitter: Shu-Ching Ou

Submission Date: Nov. 30, 2018, 10:42 a.m.

Version: 1

Publication Details
Malito E;Faleri A;Lo Surdo P;Veggi D;Maruggi G;Grassi E;Cartocci E;Bertoldi I;Genovese A;Santini L;Romagnoli G;Borgogni E;Brier S;Lo Passo C;Domina M;Castellino F;Felici F;van der Veen S;Johnson S;Lea SM;Tang CM;Pizza M;Savino S;Norais N;Rappuoli R;Bottomley MJ;Masignani V,Proc Natl Acad Sci U S A (2013) Defining a protective epitope on factor H binding protein, a key meningococcal virulence factor and vaccine antigen. PMID:23396847
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1QGC 2000-01-26 STRUCTURE OF THE COMPLEX OF A FAB FRAGMENT OF A NEUTRALIZING ANTIBODY WITH FOOT AND MOUTH DISEASE VIRUS
2AJV 2006-02-14 1.5 Crystal Structure of Cocaine catalytic Antibody 7A1 Fab' in Complex with Cocaine
2AJU 2006-02-14 1.5 Cyrstal structure of cocaine catalytic antibody 7A1 Fab'
1RUR 2004-03-02 1.5 Crystal Structure (I) of native Diels-Alder antibody 13G5 Fab at pH 8.0 with a data set collected at SSRL beamline 9-1.
2AJS 2006-02-14 1.7 Crystal structure of cocaine catalytic antibody 7A1 Fab' in complex with heptaethylene glycol
1SEQ 2005-03-01 1.78 Fab MNAC13
1Q9O 2004-01-27 1.79 S45-18 Fab Unliganded
1DQQ 2000-01-19 1.8 CRYSTAL STRUCTURE OF ANTI-LYSOZYME ANTIBODY HYHEL-63
1KCV 2002-05-11 1.8 Crystal structure of antibody pc282
1NBY 2003-04-01 1.8 Crystal Structure of HyHEL-63 complexed with HEL mutant K96A
1NBZ 2003-04-01 1.85 Crystal Structure of HyHEL-63 complexed with HEL mutant K97A
2AJX 2006-02-14 1.85 Crystal Structure of Cocaine catalytic Antibody 7A1 Fab' in Complex with Transition State Analog
2AK1 2006-02-14 1.85 Crystal Structure of Cocaine catalytic Antibody 7A1 Fab' in Complex with benzoic acid
1RUQ 2004-03-02 1.86 Crystal Structure (H) of u.v.-irradiated Diels-Alder antibody 13G5 Fab at pH 8.0 with a data set collected in house.
1ORS 2003-05-06 1.9 X-ray structure of the KvAP potassium channel voltage sensor in complex with an Fab
6FAB 1993-01-15 1.9 THREE-DIMENSIONAL STRUCTURE OF MURINE ANTI-P-AZOPHENYLARSONATE FAB 36-71. 1. X-RAY CRYSTALLOGRAPHY, SITE-DIRECTED MUTAGENESIS, AND MODELING OF THE COMPLEX WITH HAPTEN
1E4X 2001-07-12 1.9 crossreactive binding of a circularized peptide to an anti-TGFalpha antibody Fab-fragment
1NDG 2003-06-03 1.9 Crystal structure of Fab fragment of antibody HyHEL-8 complexed with its antigen lysozyme
1R3J 2003-11-25 1.9 potassium channel KcsA-Fab complex in high concentration of Tl+
1OSP 1997-04-21 1.95 CRYSTAL STRUCTURE OF OUTER SURFACE PROTEIN A OF BORRELIA BURGDORFERI COMPLEXED WITH A MURINE MONOCLONAL ANTIBODY FAB
1E4W 2001-07-12 1.95 crossreactive binding of a circularized peptide to an anti-TGFalpha antibody Fab-fragment
1GPO 1998-04-01 1.95 CRYSTAL STRUCTURE OF THE RATIONALLY DESIGNED ANTIBODY M41 AS A FAB FRAGMENT
1A3L 1999-02-16 1.95 CATALYSIS OF A DISFAVORED REACTION: AN ANTIBODY EXO DIELS-ALDERASE-TSA-INHIBITOR COMPLEX AT 1.95 A RESOLUTION
1FNS 2000-10-18 2.0 CRYSTAL STRUCTURE OF THE VON WILLEBRAND FACTOR (VWF) A1 DOMAIN I546V MUTANT IN COMPLEX WITH THE FUNCTION BLOCKING FAB NMC4
1DQJ 2000-01-19 2.0 CRYSTAL STRUCTURE OF THE ANTI-LYSOZYME ANTIBODY HYHEL-63 COMPLEXED WITH HEN EGG WHITE LYSOZYME
1NBV 1994-01-31 2.0 AN AUTOANTIBODY TO SINGLE-STRANDED DNA: COMPARISON OF THE THREE-DIMENSIONAL STRUCTURES OF THE UNLIGANDED FAB AND A DEOXYNUCLEOTIDE-FAB COMPLEX
1P2C 2004-02-17 2.0 crystal structure analysis of an anti-lysozyme antibody
25C8 1999-03-23 2.0 CATALYTIC ANTIBODY 5C8, FAB-HAPTEN COMPLEX
1UB5 2004-04-20 2.0 Crystal structure of Antibody 19G2 with hapten at 100K
2Q76 2008-03-18 2.0 Mouse anti-hen egg white lysozyme antibody F10.6.6 Fab fragment
35C8 1999-03-23 2.0 CATALYTIC ANTIBODY 5C8, FAB-INHIBITOR COMPLEX
1XGU 2005-09-06 2.1 Structure for antibody HyHEL-63 Y33F mutant complexed with hen egg lysozyme
2AJY 2006-02-14 2.1 Crystal Structure of Cocaine catalytic Antibody 7A1 Fab' in Complex with ecgonine methyl ester and benzoic acid
1DQM 2000-01-19 2.1 CRYSTAL STRUCTURE OF ANTI-LYSOZYME ANTIBODY
1MH5 2003-09-23 2.1 The Structure Of The Complex Of The Fab Fragment Of The Esterolytic Antibody MS6-164 and A Transition-State Analog
1MLB 1995-06-03 2.1 MONOCLONAL ANTIBODY FAB D44.1 RAISED AGAINST CHICKEN EGG-WHITE LYSOZYME
1NDM 2003-06-03 2.1 Crystal structure of Fab fragment of antibody HyHEL-26 complexed with lysozyme
1UB6 2004-04-20 2.12 Crystal structure of Antibody 19G2 with sera ligand
2RGS 2008-10-07 2.13 FC-fragment of monoclonal antibody IGG2B from Mus musculus
1KCU 2002-05-11 2.2 CRYSTAL STRUCTURE OF ANTIBODY PC287
1UWX 2005-06-15 2.2 P1.2 serosubtype antigen derived from N. meningitidis PorA in complex with Fab fragment
2CK0 2003-08-05 2.2 ANTI-ANTI-IDIOTYPIC ANTIBODY AGAINST HUMAN ANGIOTENSIN II, COMPLEX WITH A SYNTHETIC CYCLIC PEPTIDE
1CT8 1999-11-10 2.2 CATALYTIC ANTIBODY 7C8 COMPLEX
1EMT 2000-11-01 2.25 FAB ANTIBODY FRAGMENT OF AN C60 ANTIFULLERENE ANTIBODY
2H8P 2006-09-12 2.25 Structure of a K channel with an amide to ester substitution in the selectivity filter
1A0Q 1999-03-02 2.3 29G11 COMPLEXED WITH PHENYL [1-(1-N-SUCCINYLAMINO)PENTYL] PHOSPHONATE
2AJZ 2006-02-14 2.3 Crystal Structure of Cocaine catalytic Antibody 7A1 Fab' in Complex with ecgonine methyl ester
1K4D 2001-11-14 2.3 Potassium Channel KcsA-Fab complex in low concentration of K+
4KVC 2013-07-24 2.31 2H2 Fab fragment of immature Dengue virus
1R3I 2003-11-25 2.4 potassium channel KcsA-Fab complex in Rb+
1EHL 2001-02-21 2.4 64M-2 ANTIBODY FAB COMPLEXED WITH D(5HT)(6-4)T
1R3L 2003-11-25 2.41 potassium channel KcsA-Fab complex in Cs+
1FL3 2000-11-01 2.45 CRYSTAL STRUCTURE OF THE BLUE FLUORESCENT ANTIBODY (19G2) IN COMPLEX WITH STILBENE HAPTEN AT 277K
1MAM 1993-10-31 2.45 CRYSTAL STRUCTURE TO 2.45 A RESOLUTION OF A MONOCLONAL FAB SPECIFIC FOR THE BRUCELLA A CELL WALL POLYSACCHARIDE ANTIGEN
1KCS 2002-05-11 2.5 CRYSTAL STRUCTURE OF ANTIBODY PC282 IN COMPLEX WITH PS1 PEPTIDE
1FDL 1991-10-15 2.5 CRYSTALLOGRAPHIC REFINEMENT OF THE THREE-DIMENSIONAL STRUCTURE OF THE FAB D1.3-LYSOZYME COMPLEX AT 2.5-ANGSTROMS RESOLUTION
1CIC 1999-04-19 2.5 IDIOTOPE-ANTI-IDIOTOPE FAB-FAB COMPLEX; D1.3-E225
1FGN 1998-02-25 2.5 MONOCLONAL MURINE ANTIBODY 5G9-ANTI-HUMAN TISSUE FACTOR
1KC5 2002-07-24 2.5 CRYSTAL STRUCTURE OF ANTIBODY PC287 IN COMPLEX WITH PS1 PEPTIDE
1CK0 2003-08-05 2.5 ANTI-ANTI-IDIOTYPIC ANTIBODY AGAINST HUMAN ANGIOTENSIN II, UNLIGANDED FORM
1RIH 2004-01-13 2.5 Crystal Structure of Fab 14F7, a unique anti-tumor antibody specific for N-glycolyl GM3
15C8 1999-03-23 2.5 CATALYTIC ANTIBODY 5C8, FREE FAB
1KB5 1998-04-08 2.5 MURINE T-CELL RECEPTOR VARIABLE DOMAIN/FAB COMPLEX
1MLC 1995-06-03 2.5 MONOCLONAL ANTIBODY FAB D44.1 RAISED AGAINST CHICKEN EGG-WHITE LYSOZYME COMPLEXED WITH LYSOZYME
1OTS 2003-04-15 2.51 Structure of the Escherichia coli ClC Chloride channel and Fab Complex
1HI6 2001-02-08 2.55 ANTI-P24 (HIV-1) FAB FRAGMENT CB41 COMPLEXED WITH A PEPTIDE
1HH6 2001-01-26 2.6 ANTI-P24 (HIV-1) FAB FRAGMENT CB41 COMPLEXED WITH A PEPTIDE
1C12 1999-08-14 2.6 INSIGHT IN ODORANT PERCEPTION: THE CRYSTAL STRUCTURE AND BINDING CHARACTERISTICS OF ANTIBODY FRAGMENTS DIRECTED AGAINST THE MUSK ODORANT TRASEOLIDE
1MF2 1997-12-31 2.6 ANTI HIV1 PROTEASE FAB COMPLEX
1ETZ 2000-10-18 2.6 THE THREE-DIMENSIONAL STRUCTURE OF AN ANTI-SWEETENER FAB, NC10.14, SHOWS THE EXTENT OF STRUCTURAL DIVERSITY IN ANTIGEN RECOGNITION BY IMMUNOGLOBULINS
1CFN 1999-03-30 2.65 ANTI-P24 (HIV-1) FAB FRAGMENT CB41 COMPLEXED WITH AN EPITOPE-RELATED PEPTIDE
1CBV 1994-01-31 2.66 AN AUTOANTIBODY TO SINGLE-STRANDED DNA: COMPARISON OF THE THREE-DIMENSIONAL STRUCTURES OF THE UNLIGANDED FAB AND A DEOXYNUCLEOTIDE-FAB COMPLEX
1FJ1 2000-10-11 2.68 LYME DISEASE ANTIGEN OSPA IN COMPLEX WITH NEUTRALIZING ANTIBODY FAB LA-2
1FAI 1992-10-15 2.7 THREE-DIMENSIONAL STRUCTURE OF TWO CRYSTAL FORMS OF FAB R19.9, FROM A MONOCLONAL ANTI-ARSONATE ANTIBODY
1HH9 2001-01-12 2.7 ANTI-P24 (HIV-1) FAB FRAGMENT CB41 COMPLEXED WITH A PEPTIDE
1CF8 1999-07-28 2.7 Convergence of catalytic antibody and terpene cyclase mechanisms: polyene cyclization directed by carbocation-pi interactions
5UBX 2017-09-20 2.7 Crystal structure of a mutant mIgG2b Fc heterodimer in complex with Protein A peptide analog Z34C
1HQ4 2003-06-03 2.7 STRUCTURE OF NATIVE CATALYTIC ANTIBODY HA5-19A4
1CFS 1999-03-31 2.75 ANTI-P24 (HIV-1) FAB FRAGMENT CB41 COMPLEXED WITH AN EPITOPE-UNRELATED PEPTIDE
2HG5 2006-09-12 2.75 Cs+ complex of a K channel with an amide to ester substitution in the selectivity filter
1AI1 1997-05-15 2.8 HIV-1 V3 LOOP MIMIC
1FRG 1994-05-31 2.8 CRYSTAL STRUCTURE, SEQUENCE, AND EPITOPE MAPPING OF A PEPTIDE COMPLEX OF AN ANTI-INFLUENZA HA PEPTIDE ANTIBODY FAB 26(SLASH)9: FINE-TUNING ANTIBODY SPECIFICITY
4QNP 2015-02-11 2.8 Crystal structure of the 2009 pandemic H1N1 influenza virus neuraminidase with a neutralizing antibody
1JRH 1998-03-25 2.8 COMPLEX (ANTIBODY/ANTIGEN)
1PSK 1997-09-04 2.8 THE CRYSTAL STRUCTURE OF AN FAB FRAGMENT THAT BINDS TO THE MELANOMA-ASSOCIATED GD2 GANGLIOSIDE
2F19 1992-10-15 2.8 THREE-DIMENSIONAL STRUCTURE OF TWO CRYSTAL FORMS OF FAB R19.9, FROM A MONOCLONAL ANTI-ARSONATE ANTIBODY
1R3K 2003-11-25 2.8 potassium channel KcsA-Fab complex in low concentration of Tl+
1CFT 1999-03-31 2.8 ANTI-P24 (HIV-1) FAB FRAGMENT CB41 COMPLEXED WITH AN EPITOPE-UNRELATED D-PEPTIDE
1CFQ 1999-03-30 2.8 ANTI-P24 (HIV-1) FAB FRAGMENT CB41
4KK8 2013-08-21 2.86 Structure of the E148Q mutant of CLC-ec1 deltaNC construct in 100mM fluoride
1FSK 2000-10-02 2.9 COMPLEX FORMATION BETWEEN A FAB FRAGMENT OF A MONOCLONAL IGG ANTIBODY AND THE MAJOR ALLERGEN FROM BIRCH POLLEN BET V 1
1AIF 1997-02-01 2.9 ANTI-IDIOTYPIC FAB 409.5.3 (IGG2A) FAB FROM MOUSE
1IAI 1996-03-08 2.9 IDIOTYPE-ANTI-IDIOTYPE FAB COMPLEX
3EJZ 2009-02-17 2.9 Structure of E203V mutant E.coli Cl-/H+ exchanger, CLC-ec1
1KCR 2002-05-11 2.9 CRYSTAL STRUCTURE OF ANTIBODY PC283 IN COMPLEX WITH PS1 PEPTIDE
1FBI 1995-02-27 3.0 CRYSTAL STRUCTURE OF A CROSS-REACTION COMPLEX BETWEEN FAB F9.13.7 AND GUINEA-FOWL LYSOZYME
1OTT 2003-04-15 3.0 Structure of the Escherichia coli ClC Chloride channel E148A mutant and Fab Complex
3FB6 2010-05-19 3.0 KcsA Potassium channel in the partially open state with 16 A opening at T112
3CK0 2003-08-05 3.0 ANTI-ANTI-IDIOTYPIC ANTIBODY AGAINST HUMAN ANGIOTENSIN II, COMPLEX WITH HUMAN ANGIOTENSIN II
1AHW 1998-02-25 3.0 A COMPLEX OF EXTRACELLULAR DOMAIN OF TISSUE FACTOR WITH AN INHIBITORY FAB (5G9)
1N5Y 2003-01-28 3.1 HIV-1 Reverse Transcriptase Crosslinked to Post-Translocation AZTMP-Terminated DNA (Complex P)
4KK5 2013-08-21 3.17 Structure of the CLC-ec1 deltaNC construct in 20mM fluoride and 20mM bromide
1KNO 1996-01-29 3.2 CRYSTAL STRUCTURE OF THE COMPLEX OF A CATALYTIC ANTIBODY FAB WITH A TRANSITION STATE ANALOG: STRUCTURAL SIMILARITIES IN ESTERASE-LIKE ABZYMES
1IGY 1998-04-15 3.2 STRUCTURE OF IMMUNOGLOBULIN
3F7V 2010-05-19 3.2 KcsA Potassium channel in the open-inactivated state with 23 A opening at T112
1ORQ 2003-05-06 3.2 X-ray structure of a voltage-dependent potassium channel in complex with an Fab
1OTU 2003-04-15 3.3 Structure of the Escherichia coli ClC Chloride channel E148Q mutant and Fab Complex
3F7Y 2010-05-19 3.4 KcsA Potassium channel in the partially open state with 17 A opening at T112
1KEN 2002-04-24 3.5 INFLUENZA VIRUS HEMAGGLUTININ COMPLEXED WITH AN ANTIBODY THAT PREVENTS THE HEMAGGLUTININ LOW PH FUSOGENIC TRANSITION

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 L fHbp-Fab 12C1 P01837 IGKC_MOUSE
100.0 H fHbp-Fab 12C1 P01867 IGG2B_MOUSE