Research Capabilities
Protein and Peptide Markers of Infection
Using mass spectrometry-based discovery tools, the secreted protein marker signature indicative of Francisella spp. infection will be determined for samples from both the 3-D cell culture system and murine model.
PNNL will use this technique to address the protein and peptide markers of infection, where the secreted protein marker signature indicative of Francisella spp. infection will be determined from the PNNL three-dimensional (3-D) cell culture system and collaborator murine model using mass spectrometry-based discovery tools. This project will employ selective and semi-selective methods to target classes of protein and peptide markers of infection in our model system, Francisella spp. The goal is to determine the protein signature related to infection relative to uninfected controls. Samples from 3-D cell culture or mouse model bronchoalveolar lavage fluid will also be available as parallel sample sets. Protein and peptide markers will be analyzed by liquid chromatography and tandem mass spectrometry after a targeted extraction and cleanup. Protein marker discovery will use a marker display and comparison approach as well as a bottom-up proteomics protein identification approach. The latter approach is made possible by the use of host systems with fully sequenced genomic databases. The resulting protein marker data will be integrated with metabolite data generated by the novel volatile organic signature project. This integrated effort will provide markers that will be validated using affinity-based assays, which will later be developed into field-deployable marker detection platforms.