Early markers of kidney transplant rejection:
Proteomic workflows for discovery and the development of non-invasive, targeted quantitation assays
Amy Zumwalt1, David Sarracino2, Waichi Wong3, Emmanuel Zorn3, Bryan Krastins2, Michael Athanas4, Amol Prakash2 and Mary F Lopez2
1Thermo Fisher Scientific, San Jose, CA
2BRIMS Center, Thermo Fisher Scientific, Cambridge, MA
3Renal and Transplant Units, Massachusetts Gen. Hospital, Harvard Med. School, Boston, MA
4Vast Scientific, Wayland, MA
The accurate diagnosis of renal allograft rejection currently depends on a biopsy. Transplant medicine would benefit greatly from the availability of non-invasive tests for early detection of rejection as well as immunosuppressive drug therapeutic monitoring. This presentation describes the use of an LTQ Orbitrap XL hybrid mass spectrometer specifically geared toward the identification of differentially expressed proteins and endogenous peptides in urine. A panel of the protein or peptide biomarkers identified in the high-resolution discovery workflow was subsequently quantified in targeted selective reaction monitoring (SRM) assays on a TSQ Quantum Ultra triple quadrupole mass spectrometer. This workflow was used to analyze urine samples from normal, early rejection and acute humoral rejection transplant patients.
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