The PSWRCE Protein and PCR MS Core will collaborate on the following projects:
Specific Aim i - Proteomics Analyses for Burkholderia, Coccidioides, and Reservoir Vaccines 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 Specific Aim 2 - Development of Diagnostic Assays ? Use biomarker IDs of Sp.
Aim i 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 Specific Aim 3 - Ibis TSOOO PCR-MS Biosensor Applied to Reservoir Vaccines, Burkholderia &Coccidioides ? PCR MS (on mitochondrial DNA) for ID of vertebrate host on which vector has fed (e.g., tick nymph fed on mule deer) 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 Specific Aim 4 - Characterization of Protein Complexes (uncompensated synergistic activity) ? Structurally characterize large protein-protein complexes (e.g., pili involved in virulence, assemblies of bacteriophage tail fiber proteins useful for biosensors) using a one-of-a-kind modified QTOF MS The antigen discovery coupled to support of diagnostic assay development, PCR-MS, bioaffinity MS, and structural characterization of large protein-protein complexes are unique capabilities of the core.
Collaborations of the Core with RCE projects will allow: identification of proteins that will be used in diagnostic tests or as vaccine candidates;determination of protein expression differences between virulent and non-virulent strains of organisms;identification of vertebrate hosts of tick vectors, allowing disease transmission to be better tracked;pathogens to be rapidly characterized by PCR MS;and characterization of large protein complexes important in virulence or diagnostic design.
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