Antibody-based screen for ionizing radiation-dependent changes in the Mammalian proteome for use in biodosimetry.

In an effort to identify proteomic changes that may be useful for radiation biodosimetry, human cells of hematological origin were treated with ionizing radiation or mock-irradiated and then harvested at different times after treatment. Protein lysates were generated from these cells and evaluated by Western blotting using a panel of 301 commercially available antibodies targeting 161 unique proteins. From this screen, we identified 55 ionizing radiation-responsive proteins, including 14 proteins not previously reported to be radiation-responsive at the protein level. The data from this large-scale screen have been assembled into a public website ( http://labs.fhcrc.org/paulovich/biodose_index.html ) that may be of value to the radiation community both as a source of putative biomarkers for biodosimetry and also as a source of validation data on commercially available antibodies that detect radiation-responsive proteins. Using a panel of candidate radiation biomarkers in human cell lines, we demonstrate the feasibility of assembling a complementary panel of radiation-responsive proteins. Furthermore, we demonstrate the feasibility of using blood cell-based proteomic changes for biodosimetry by demonstrating detection of protein changes in circulating cells after total-body irradiation in a canine model.