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Multimodal Mass Spectrometry Identifies a Conserved Protective Epitope in S. pyogenes Streptolysin O

Tang, Di LU orcid ; Gueto-Tettay, Carlos LU ; Hjortswang, Elisabeth LU orcid ; Ströbaek, Joel LU ; Ekström, Simon LU ; Happonen, Lotta LU ; Malmström, Lars LU and Malmström, Johan LU orcid (2024) In Analytical Chemistry
Abstract
An important element of antibody-guided vaccine design is the use of neutralizing or opsonic monoclonal antibodies to define protective epitopes in their native three-dimensional conformation. Here, we demonstrate a multimodal mass spectrometry-based strategy for in-depth characterization of antigen–antibody complexes to enable the identification of protective epitopes using the cytolytic exotoxin Streptolysin O (SLO) from Streptococcus pyogenes as a showcase. We first discovered a monoclonal antibody with an undisclosed sequence capable of neutralizing SLO-mediated cytolysis. The amino acid sequence of both the antibody light and the heavy chain was determined using mass-spectrometry-based de novo sequencing, followed by... (More)
An important element of antibody-guided vaccine design is the use of neutralizing or opsonic monoclonal antibodies to define protective epitopes in their native three-dimensional conformation. Here, we demonstrate a multimodal mass spectrometry-based strategy for in-depth characterization of antigen–antibody complexes to enable the identification of protective epitopes using the cytolytic exotoxin Streptolysin O (SLO) from Streptococcus pyogenes as a showcase. We first discovered a monoclonal antibody with an undisclosed sequence capable of neutralizing SLO-mediated cytolysis. The amino acid sequence of both the antibody light and the heavy chain was determined using mass-spectrometry-based de novo sequencing, followed by chemical cross-linking mass spectrometry to generate distance constraints between the antibody fragment antigen-binding region and SLO. Subsequent integrative computational modeling revealed a discontinuous epitope located in domain 3 of SLO that was experimentally validated by hydrogen–deuterium exchange mass spectrometry and reverse engineering of the targeted epitope. The results show that the antibody inhibits SLO-mediated cytolysis by binding to a discontinuous epitope in domain 3, likely preventing oligomerization and subsequent secondary structure transitions critical for pore-formation. The epitope is highly conserved across >98% of the characterized S. pyogenes isolates, making it an attractive target for antibody-based therapy and vaccine design against severe streptococcal infections. (Less)
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author
; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
epub
subject
in
Analytical Chemistry
publisher
The American Chemical Society (ACS)
external identifiers
  • pmid:38701337
ISSN
1520-6882
DOI
10.1021/acs.analchem.4c00596
project
Proof-of-concept design of multivalent nanoparticle vaccines against antibiotic-resistant bacteria
Properties of Protective Antibody Responses against Bacterial Pathogens
language
English
LU publication?
yes
id
5d070d7b-46b1-4bdc-b1a2-62b1a026cd78
date added to LUP
2024-05-06 16:39:46
date last changed
2024-05-08 03:00:03
@article{5d070d7b-46b1-4bdc-b1a2-62b1a026cd78,
  abstract     = {{An important element of antibody-guided vaccine design is the use of neutralizing or opsonic monoclonal antibodies to define protective epitopes in their native three-dimensional conformation. Here, we demonstrate a multimodal mass spectrometry-based strategy for in-depth characterization of antigen–antibody complexes to enable the identification of protective epitopes using the cytolytic exotoxin Streptolysin O (SLO) from <i>Streptococcus pyogenes</i> as a showcase. We first discovered a monoclonal antibody with an undisclosed sequence capable of neutralizing SLO-mediated cytolysis. The amino acid sequence of both the antibody light and the heavy chain was determined using mass-spectrometry-based <i>de novo</i> sequencing, followed by chemical cross-linking mass spectrometry to generate distance constraints between the antibody fragment antigen-binding region and SLO. Subsequent integrative computational modeling revealed a discontinuous epitope located in domain 3 of SLO that was experimentally validated by hydrogen–deuterium exchange mass spectrometry and reverse engineering of the targeted epitope. The results show that the antibody inhibits SLO-mediated cytolysis by binding to a discontinuous epitope in domain 3, likely preventing oligomerization and subsequent secondary structure transitions critical for pore-formation. The epitope is highly conserved across &gt;98% of the characterized <i>S. pyogenes</i> isolates, making it an attractive target for antibody-based therapy and vaccine design against severe streptococcal infections.}},
  author       = {{Tang, Di and Gueto-Tettay, Carlos and Hjortswang, Elisabeth and Ströbaek, Joel and Ekström, Simon and Happonen, Lotta and Malmström, Lars and Malmström, Johan}},
  issn         = {{1520-6882}},
  language     = {{eng}},
  month        = {{05}},
  publisher    = {{The American Chemical Society (ACS)}},
  series       = {{Analytical Chemistry}},
  title        = {{Multimodal Mass Spectrometry Identifies a Conserved Protective Epitope in <i>S. pyogenes</i> Streptolysin O}},
  url          = {{http://dx.doi.org/10.1021/acs.analchem.4c00596}},
  doi          = {{10.1021/acs.analchem.4c00596}},
  year         = {{2024}},
}