Biomarkers of Exposure to Toxic Substances Volume 7: Identification of Potential Serum Protein Biomarkers Indicative of Low Level Kidney Degradation in Response to Toxin Exposures
Final rept. 1 oct 2005-30 Apr 2009
AIR FORCE RESEARCH LAB WRIGHT-PATTERSON AFB OH HUMAN EFFECTIVENESS DIRECTORATE
Pagination or Media Count:
Potential serum biomarkers to subclinical nephrotoxin exposures were evaluated based on differential protein expression between control and dosed samples in rat serum. Proteins of interest demonstrated up-regulation at a minimum 1.5 fold increase in protein concentration control versus dosed sample. In order to identify common biomarkers of kidney decrement, three nephrotoxicants were chosen D-serine, Puromycin, an antibiotic that degrades and Bromoethyl Amine BEA. Rats were dosed with individual nephrotoxins at levels previously determined, via hisopathology, to initiate low level renal damage. Blood serum was collected at pre-dose, 24 hours post dose, and terminal sacrifice for puromycin and BEA, and at 12 and 24 hours post dose from D-serine. Method development of the immunodepletion step demonstrated that the Proteomet Lab IgY-R7 column worked substantially better than the Agilent MARS HPLC and Spin columns in a rat sera matrix. The collected serum samples were immunodepleted using the IgY-R7 column, separated using 2D Difference in Gel Electrophoresis DIGE, and screened for potential up-regulated biomarkers based on differences in fluorescent intensity via computer software analyses. Serum peptides were identified by MALDI-TOFTOF mass spectroscopy and MASCOT database alignments. This study identified 32 serum proteins that were up-regulated in response to D-serine exposures.
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