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

Publication Type:

Journal Article

Source:

Radiation research, Volume 171, Issue 5, p.549-61 (2009)

Keywords:

2009, Animals, Antibodies, Blotting, Western, Cell Line, Clinical Research Division, Dogs, Humans, PHOSPHORYLATION, Proteome, Radiation Dosage, Research Design

Abstract:

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.