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With a newly-obtained fiery red blood meal visible through her transparent abdomen, the now heavy female Aedes aegypti mosquito takes flight. Clostridium botulinum growing on egg yolk agar showing the lipase reaction.An atypical enlarged lymphocyte found in the blood smear from a HPS patient. (a) An atypical enlarged lymphocyte found in the blood smear from a HPS patient. (b)Burkholderia psudeomallei grown on sheep blood agar for 24 hours.

MASS SPECTROMETRY AND PROTEOMICS CORE

 


Protein and PCR-MS Core

Vicki Wysocki, University of Arizona

Abstract:
The specific aims of the Protein and PCR-MS Core are:

  1. Core mass spectrometry and proteomics services offered on a fee-per-sample basis to any RCE project or Region IX investigator who needs the service.
  2. Technology development that will address future analysis needs of RCE investigators.
  3. Targeted collaborative projects directly supporting projects in the RCE.

Specific aims of the Mass Spectrometry/Proteomics Core efforts are:

1. Proteomic analyses for Burkholderia, Coccidioides, and bacterial zoonoses projects:

  • Identify protein biomarkers for use as diagnostic or vaccine candidates (from serum, urine, and lung homogenates, with depletion of abundant mouse/human proteins as necessary)
  • Perform comparative proteomics on pre-fractionated samples (e.g., extracellular proteins or cell wall proteins of two different strains of an organism, or wild type vs. mutant)
  • Build “blood meal” protein library to use to identify host vertebrate proteins in tick nymphs

2. Development of Diagnostic Assays

  • Use biomarker IDs of Specific Aim 1 to develop bioaffinity MS for pathogen identification
  • Use MS to guide development of targeted ELISA or other fluorescence assays (characterize expressed protein antigens, antibodies, and labeled antibodies by MS throughout development of assay)
  • Compare antibodies vs. small ligands for capture of antigens in bioaffinity MS

3. Ibis T5000 PCR-MS Biosensor applied to bacterial zoonoses, Burkholderia & Coccidioides

  • PCR MS (on mitochondrial DNA) for ID of vertebrate host on which vector has fed and ID of tick-borne pathogenic organism present
  • PCR MS for rapid ID and strain typing of pathogens, as well as confirmation of the presence of toxin genes, antibiotic resistance genes, selected virulence factor genes

4. Characterization of protein complexes using a one-of-a-kind modified QTOF MS, PCR-MS or bioaffinity MS

http://www.chem.arizona.edu/facilities/msf/index.html
 

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