skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: High Resolution Mapping of Bactericidal Monoclonal Antibody Binding Epitopes on Staphylococcus aureus Antigen MntC

Abstract

The Staphylococcus aureus manganese transporter protein MntC is under investigation as a component of a prophylactic S.aureus vaccine. Passive immunization with monoclonal antibodies mAB 305-78-7 and mAB 305-101-8 produced using MntC was shown to significantly reduce S. aureus burden in an infant rat model of infection. Earlier interference mapping suggested that a total of 23 monoclonal antibodies generated against MntC could be subdivided into three interference groups, representing three independent immunogenic regions. In the current work binding epitopes for selected representatives of each of these interference groups (mAB 305-72-5 – group 1, mAB 305-78-7 – group 2, and mAB 305-101-8 – group 3) were mapped using Hydrogen-Deuterium Exchange Mass Spectrometry (DXMS). All of the identified epitopes are discontinuous, with binding surface formed by structural elements that are separated within the primary sequence of the protein but adjacent in the context of the three-dimensional structure. The approach was validated by co-crystallizing the Fab fragment of one of the antibodies (mAB 305-78-7) with MntC and solving the three-dimensional structure of the complex. X-ray results themselves and localization of the mAB 305-78-7 epitope were further validated using antibody binding experiments with MntC variants containing substitutions of key amino acid residues. These results providedmore » insight into the antigenic properties of MntC and how these properties may play a role in protecting the hostagainst S. aureus infection by preventing the capture and transport of Mn2+, a key element that the pathogen uses to evade host immunity.« less

Authors:
 [1];  [2];  [2];  [3];  [1];  [1];  [1];  [1];  [1];  [4]
  1. Pfizer Vaccine Research and Development, Pearl River, NY (United States)
  2. Pfizer Protein Engineering and Production, Cambridge, MA (United States)
  3. Univ. of California at San Diego, La Jolla, CA (United States)
  4. National Jewish Health, Denver, CO (United States)
Publication Date:
Research Org.:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1328023
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
PLoS Pathogens
Additional Journal Information:
Journal Volume: 12; Journal Issue: 9; Journal ID: ISSN 1553-7374
Publisher:
Public Library of Science
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; deuterium; monoclonal antibodies; antibodies; epitope mapping; X-ray crystallography; enzyme-linked immunoassays; amino acid substitution; Staphylococcus aureus

Citation Formats

Gribenko, Alexey V., Parris, Kevin, Mosyak, Lidia, Li, Sheng, Handke, Luke, Hawkins, Julio C., Severina, Elena, Matsuka, Yury V., Anderson, Annaliesa S., and Zhang, Gongyi. High Resolution Mapping of Bactericidal Monoclonal Antibody Binding Epitopes on Staphylococcus aureus Antigen MntC. United States: N. p., 2016. Web. doi:10.1371/journal.ppat.1005908.
Gribenko, Alexey V., Parris, Kevin, Mosyak, Lidia, Li, Sheng, Handke, Luke, Hawkins, Julio C., Severina, Elena, Matsuka, Yury V., Anderson, Annaliesa S., & Zhang, Gongyi. High Resolution Mapping of Bactericidal Monoclonal Antibody Binding Epitopes on Staphylococcus aureus Antigen MntC. United States. https://doi.org/10.1371/journal.ppat.1005908
Gribenko, Alexey V., Parris, Kevin, Mosyak, Lidia, Li, Sheng, Handke, Luke, Hawkins, Julio C., Severina, Elena, Matsuka, Yury V., Anderson, Annaliesa S., and Zhang, Gongyi. 2016. "High Resolution Mapping of Bactericidal Monoclonal Antibody Binding Epitopes on Staphylococcus aureus Antigen MntC". United States. https://doi.org/10.1371/journal.ppat.1005908. https://www.osti.gov/servlets/purl/1328023.
@article{osti_1328023,
title = {High Resolution Mapping of Bactericidal Monoclonal Antibody Binding Epitopes on Staphylococcus aureus Antigen MntC},
author = {Gribenko, Alexey V. and Parris, Kevin and Mosyak, Lidia and Li, Sheng and Handke, Luke and Hawkins, Julio C. and Severina, Elena and Matsuka, Yury V. and Anderson, Annaliesa S. and Zhang, Gongyi},
abstractNote = {The Staphylococcus aureus manganese transporter protein MntC is under investigation as a component of a prophylactic S.aureus vaccine. Passive immunization with monoclonal antibodies mAB 305-78-7 and mAB 305-101-8 produced using MntC was shown to significantly reduce S. aureus burden in an infant rat model of infection. Earlier interference mapping suggested that a total of 23 monoclonal antibodies generated against MntC could be subdivided into three interference groups, representing three independent immunogenic regions. In the current work binding epitopes for selected representatives of each of these interference groups (mAB 305-72-5 – group 1, mAB 305-78-7 – group 2, and mAB 305-101-8 – group 3) were mapped using Hydrogen-Deuterium Exchange Mass Spectrometry (DXMS). All of the identified epitopes are discontinuous, with binding surface formed by structural elements that are separated within the primary sequence of the protein but adjacent in the context of the three-dimensional structure. The approach was validated by co-crystallizing the Fab fragment of one of the antibodies (mAB 305-78-7) with MntC and solving the three-dimensional structure of the complex. X-ray results themselves and localization of the mAB 305-78-7 epitope were further validated using antibody binding experiments with MntC variants containing substitutions of key amino acid residues. These results provided insight into the antigenic properties of MntC and how these properties may play a role in protecting the hostagainst S. aureus infection by preventing the capture and transport of Mn2+, a key element that the pathogen uses to evade host immunity.},
doi = {10.1371/journal.ppat.1005908},
url = {https://www.osti.gov/biblio/1328023}, journal = {PLoS Pathogens},
issn = {1553-7374},
number = 9,
volume = 12,
place = {United States},
year = {Fri Sep 30 00:00:00 EDT 2016},
month = {Fri Sep 30 00:00:00 EDT 2016}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 21 works
Citation information provided by
Web of Science

Save / Share:

Works referenced in this record:

Protein hydrogen exchange at residue resolution by proteolytic fragmentation mass spectrometry analysis
journal, September 2013


Defining a protective epitope on factor H binding protein, a key meningococcal virulence factor and vaccine antigen
journal, February 2013


Design and Characterization of Epitope-Scaffold Immunogens That Present the Motavizumab Epitope from Respiratory Syncytial Virus
journal, June 2011


Evaluation of Approaches to Monitor Staphylococcus aureus Virulence Factor Expression during Human Disease
journal, February 2015


Early Hydrophobic Collapse of α1-Antitrypsin Facilitates Formation of a Metastable State: Insights from Oxidative Labeling and Mass Spectrometry
journal, November 2012


Determination of the Mechanism of Action of Anti-FasL Antibody by Epitope Mapping and Homology Modeling
journal, August 2009


Protein structure change studied by hydrogen-deuterium exchange, functional labeling, and mass spectrometry
journal, May 2003


Epitope mapping of inhibitory antibodies targeting the C2 domain of coagulation factor VIII by hydrogen-deuterium exchange mass spectrometry
journal, December 2013


Structure-Based Design of a Fusion Glycoprotein Vaccine for Respiratory Syncytial Virus
journal, October 2013


IL-1β Epitope Mapping Using Site-Directed Mutagenesis and Hydrogen−Deuterium Exchange Mass Spectrometry Analysis
journal, August 2005


Conformational epitope mapping of Pru du 6, a major allergen from almond nut
journal, October 2013


CED: a conformational epitope database
journal, January 2006


Staphylococcus aureus Manganese Transport Protein C Is a Highly Conserved Cell Surface Protein That Elicits Protective Immunity Against S. aureus and Staphylococcus epidermidis
journal, April 2012


Folding mechanism of the metastable serpin  1-antitrypsin
journal, March 2012


Dynamics of cAPK Type IIβ Activation Revealed by Enhanced Amide H/2H Exchange Mass Spectrometry (DXMS)
journal, April 2003


Differential hydrogen/deuterium exchange mass spectrometry analysis of protein–ligand interactions
journal, February 2011


Primary structure effects on peptide group hydrogen exchange
journal, September 1993


Molecular epitope identification by limited proteolysis of an immobilized antigen-antibody complex and mass spectrometric peptide mapping.
journal, December 1990


Structural Insights into Fibrinogen Dynamics Using Amide Hydrogen/Deuterium Exchange Mass Spectrometry
journal, July 2013


Epitope mapping by amide hydrogen/deuterium exchange coupled with immobilization of antibody, on-line proteolysis, liquid chromatography and mass spectrometry
journal, March 2009


Regulation of Staphylococcus aureus MntC Expression and Its Role in Response to Oxidative Stress
journal, October 2013


Phage display: Concept, innovations, applications and future
journal, November 2010


Coot model-building tools for molecular graphics
journal, November 2004


Epitope mapping of human VWF A3 recognized by monoclonal antibody SZ-123 and SZ-125 using MALDI mass spectrometry
journal, August 2011


Emerging mass spectrometry-based approaches to probe protein–receptor interactions: Focus on overcoming physiological barriers
journal, July 2013


Three-Dimensional Structure and Biophysical Characterization of Staphylococcus aureus Cell Surface Antigen–Manganese Transporter MntC
journal, September 2013


Features and development of Coot
journal, March 2010


Structural Analysis of Bacterial ABC Transporter Inhibition by an Antibody Fragment
journal, April 2015


Structure and properties of  -synuclein and other amyloids determined at the amino acid level
journal, October 2005


What Is a B-Cell Epitope?
book, January 2009


Mapping of discontinuous conformational epitopes by amide hydrogen/deuterium exchange mass spectrometry and computational docking: EPITOPE MAPPING BY HDX-DOCK
journal, February 2012


[20] Processing of X-ray diffraction data collected in oscillation mode
book, January 1997


Rapid refinement of crystallographic protein construct definition employing enhanced hydrogen/deuterium exchange MS
journal, January 2004


Exploiting structure similarity in refinement: automated NCS and target-structure restraints in BUSTER
journal, March 2012


Phaser crystallographic software
journal, July 2007


Mapping of discontinuous conformational epitopes by amide hydrogen/deuterium exchange mass spectrometry and computational docking: EPITOPE MAPPING BY HDX-DOCK
journal, February 2012


What Is a B Cell Epitope
book, January 2019


Epitope mapping of human VWF A3 recognized by monoclonal antibody SZ-123 and SZ-125 using MALDI mass spectrometry
journal, August 2011


Emerging mass spectrometry-based approaches to probe protein–receptor interactions: Focus on overcoming physiological barriers
journal, July 2013


Phage display: Concept, innovations, applications and future
journal, November 2010


Applications of peptide arrays prepared by the SPOT-technology
journal, February 2001


Structural Insights into Fibrinogen Dynamics Using Amide Hydrogen/Deuterium Exchange Mass Spectrometry
journal, July 2013


Determination of the Mechanism of Action of Anti-FasL Antibody by Epitope Mapping and Homology Modeling
journal, August 2009


Rapid refinement of crystallographic protein construct definition employing enhanced hydrogen/deuterium exchange MS
journal, January 2004


Structure and properties of  -synuclein and other amyloids determined at the amino acid level
journal, October 2005


Folding mechanism of the metastable serpin  1-antitrypsin
journal, March 2012


Defining a protective epitope on factor H binding protein, a key meningococcal virulence factor and vaccine antigen
journal, February 2013


Protein structure change studied by hydrogen-deuterium exchange, functional labeling, and mass spectrometry
journal, May 2003


Protein hydrogen exchange at residue resolution by proteolytic fragmentation mass spectrometry analysis
journal, September 2013


Molecular epitope identification by limited proteolysis of an immobilized antigen-antibody complex and mass spectrometric peptide mapping.
journal, December 1990


Staphylococcus aureus Manganese Transport Protein C Is a Highly Conserved Cell Surface Protein That Elicits Protective Immunity Against S. aureus and Staphylococcus epidermidis
journal, April 2012


Epitope mapping of inhibitory antibodies targeting the C2 domain of coagulation factor VIII by hydrogen-deuterium exchange mass spectrometry
journal, December 2013


Evaluation of Approaches to Monitor Staphylococcus aureus Virulence Factor Expression during Human Disease
journal, February 2015


Differential hydrogen/deuterium exchange mass spectrometry analysis of protein–ligand interactions
journal, February 2011


Works referencing / citing this record:

Anti-bacterial Monoclonal Antibodies
book, January 2017


An overview of hydrogen deuterium exchange mass spectrometry (HDX-MS) in drug discovery
journal, August 2017


The development of a staphylococcus aureus four antigen vaccine for use prior to elective orthopedic surgery
journal, September 2018


High-Definition Mapping of Four Spatially Distinct Neutralizing Epitope Clusters on RiVax, a Candidate Ricin Toxin Subunit Vaccine
journal, October 2017


Two Vaccines for Staphylococcus aureus Induce a B-Cell-Mediated Immune Response
journal, August 2018


MntC-Dependent Manganese Transport Is Essential for Staphylococcus aureus Oxidative Stress Resistance and Virulence
journal, July 2018


Tracking Higher Order Protein Structure by Hydrogen-Deuterium Exchange Mass Spectrometry
journal, February 2019


A Collection of Single-Domain Antibodies that Crowd Ricin Toxin’s Active Site
journal, December 2018


Persistence of Immune Responses Through 36 Months in Healthy Adults After Vaccination With a Novel Staphylococcus aureus 4-Antigen Vaccine (SA4Ag)
journal, December 2019