A gel-free proteomic-based method for the characterization of Bordetella pertussis clinical isolates

Details

• Alternative Title:
  J Microbiol Methods
• Personal Author:
  Williamson, Yulanda M. ; Moura, Hercules ; Simmons, Kaneatra ; Whitmon, Jennifer ; Melnick, Nikkol ; Rees, Jon ; Woolfitt, Adrian ; Schieltz, David M. ; Tondella, Maria L. ; Ades, Edwin ; Sampson, Jacquelyn ; Carlone, George ; Barr, John R.
• Description:
  Bordetella pertussis (Bp) is the etiologic agent of pertussis or whooping cough, a highly contagious respiratory disease occurring primarily in infants and young children. Although vaccine preventable, pertussis cases have increased over the years leading researchers to re-evaluate vaccine control strategies. Since bacterial outer membrane proteins, comprising the surfaceome, often play roles in pathogenesis and antibody-mediated immunity, three recent Bp circulating isolates were examined using proteomics to identify any potential changes in surface protein expression. Fractions enriched for outer membrane proteins were digested with trypsin and the peptides analyzed by nano liquid chromatography-electrospray ionization-mass spectrometry (nLC-ESI-MS), followed by database analysis to elucidate the surfaceomes of our three Bp isolates. Furthermore, a less labor intensive non-gel based antibody affinity capture technology in conjunction with MS was employed to assess each Bp strains' immunogenic outer membrane proteins. This novel technique is generally applicable allowing for the identification of immunogenic surface expressed proteins on pertussis and other pathogenic bacteria.
• Subjects:
  Animals Antibodies, Bacterial Antibody-affinity Antigens, Bacterial Article Bacterial Proteins Bordetella Pertussis Child, Preschool Chromatography, Liquid Computational Biology Female Gel-free Humans Infant LC-MS/MS Membrane Mice Mice, Inbred BALB C Pertussis Protein Binding Proteome Proteomics Spectrometry, Mass, Electrospray Ionization Whooping Cough

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• Source:
  J Microbiol Methods. 90(2):119-133
• Pubmed ID:
  22537821
• Pubmed Central ID:
  PMC5687064
• Document Type:
  Journal Article
• Funding:
  CC999999/ImCDC/Intramural CDC HHSUnited States/
• Volume:
  90
• Issue:
  2
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:4f9d434e282c29b0280ca8969b99c482074b4a57bbe61e12cced2be20182a431
• Download URL:
  https://stacks.cdc.gov/view/cdc/49846/cdc_49846_DS1.pdf
• File Type:
  [PDF - 1.48 MB ]