NIH Proteomics Interest Group

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ProtIG is an NIH Special Interest Group (SIG) that organizes seminars and workshops in relevant areas of proteomics, including talks on separation and protein identification methods, determination of post-translational modifications, protein-protein interactions, and bioinformatics and data management. A monthly seminar series is held at 10am usually on the first Thursday of each month (always check the Mtgs/Seminars button on this page for these and other PROTIG announced meetings). To receive email announcements of ProtIG events, join the listserv (Join the SIG button on this page)

October ProtIG Seminar
October 1, 2015
9:30am - 10:30pm
Building 50, NIH Campus
Room 1328/1334 (Rear)
Lisa Cazares, Ph.D.
Senior Scientist, Molecular and Translational Sciences Division of US Army Research Institute of Infectious Disease (USAMRIID)

"Proteomic Analysis of Serum and Tissue for the Detection and Discovery of Early Markers of Filovirus Infection"

Ebola virus (EBOV) and Marburg virus (MARV), two species within the family Filoviridae, are among the most pathogenic human viruses known and are the causative agents of hemorrhagic fever in humans and non-human primates (NHP). A confounding issue for physicians and point of care personnel is that, the initial symptoms of filovirus are so general that they are readily mistaken for many other diseases. Our proteomic efforts at USAMRIID in the Division of Molecular and Translational Sciences are focused on the discovery of early markers of filovirus infection which can be used for diagnosis, as well as a trigger to initiate treatment with antiviral therapeutics. Using EBOV infected NHP plasma, our experimental strategy first involves inactivation of the virus using SDS buffer, followed by filter assisted sample preparation (FASP) after removal from BSL-4 containment. FASP captures the serum proteins onto a membrane where they are digested with trypsin, and the resulting peptides are labelled with reporter ion tags (TMT) to allow for the relative quantitation of protein abundance. In addition, the use of 6-plex TMT tags allows for the quantitation of proteins in 6 conditions simultaneously (i.e. pre-infection, and 5 post-infection time-points). Using LC-MS/MS on a LTQ Orbitrap Elite, we have completed a preliminary interrogation of NHP host response proteins over the course of EBOV infection in 7 NHP sample sets, each with 6 time-points. We have demonstrated for the first time that several acute phase proteins are induced systematically prior to the first detection of viremia. Comparison of the EBOV NHP host response to the response observed in 5 Burkholderia psudomallei infected NHP resulted in the discovery of common and differentially expressed proteins in these two infection types. Additional proteomic studies using EBOV infected tissue for biomarker and therapeutic target discovery are also being conducted. We have successfully employed heat fixation for the inactivation of filoviruses in infected NHP tissue. This allows for the preservation of proteins without the cross-linking issues which render formalin fixed tissue problematic for proteomic analysis. We have found heat fixed tissue suitable for many downstream proteomic and metabolomics applications including MALDI mass spectrometry imaging (MALDI-MSI) of thin tissue sections.

Seminars will be webcast online at and available on the
Proteomics Interest Group website as an archived presentation unless otherwise noted.

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This site was updated on September 26th, 2015. Please contact Renee Olano at olanol(at) with questions or suggestions.