The requirement is for analysis of RAW264.7 cells by 5 strains of Burkholderia pseudomallei at two different time points after infection. The experiments will be performed in duplicate for a total of 20 samples.
B. pseudomallei is an intracellular pathogen that can survive and replicate inside phagocytic cells. It escapes the phagosome and enters the cytosol of eukaryotic cells, where it replicates to high numbers. It is thought that B. pseudomallei secretes protein effectors while inside eukaryotic cells that enable it to replicate and resist killing, but the effectors are unknown.
USAMRIID seeks to identify bacterial effectors secreted inside RAW264.7 cells and identify the host proteins that are modulated by these effectors.
Twenty filter-sterilized protein samples will be prepared by the USAMRIID investigators and sent to the contractor for mass spectrometry analysis. The contractor should perform global proteomic profiling (including sample preparation), 16 hour MED-SAX-MS/MS on Q-Exactive Orbitrap, 4 Fractions, inclusive of sample prep, data analysis and relative quantitation profiling.
These analyses should be run on a system that utilizes nanoflow liquid chromatography coupled via electrospray ionization to a linear ion trap mass spectrometer. The system should deliver sub-femtomole sensitivity. The data should be searched using up to four search algorithms to provide maximum sequence coverage, sensitivity and specificity.
Further details are available under Solicitation Number: W81XWH14T0451. The response deadline is September 15, 2014.