Project Summary
Many NIH-funded faculties at Tulane University require state-of-art LC-MS/MS to conduct biomedical research
projects intended for clinical translation. A survey of these users revealed several features that were missing
from instruments available to Tulane faculty members, while our research on available instruments found one
set of instruments that could address these unmet needs: Orbitrap Eclipse Tribrid mass analyzer systems. The
proposed ultra-high-sensitive Orbitrap Eclipse Tribrid LC-MS/MS system possesses several unique features that
are essential for our research but are not available on our existing platform, such as single-cell proteomics,
multistage mass spectrometry (MSn) capability, increased precursor dissociation options, and Real-time Search
Synchronous Precursor Selection (SPS MS3) Tandem Mass Tag (TMT) data acquisition mode.
We have confirmed that the Orbitrap Eclipse Tribrid mass spectrometer with nano liquid chromatography,
UltiMate 3000 RSLCnano (U3000 LC) system meets or exceeds the needs of our user base and offers superior
performance relative to other similar available LC-MS/MS systems. The proposed instrument will support the
discovery of new biomarkers and the development of new methods of significant biomedical relevance, which
we expect will lead to breakthroughs in disease diagnosis, mechanism studies, and targeted therapy.
The proposed Orbitrap Eclipse Tribrid mass spectrometry with U3000 nanoLC system represents the next
generation of LC-MS/MS system. The instrument provides higher resolution and faster scan speeds, and can
perform parallel acquisition to maximize the duty cycle. Our demonstration project with Thermo Scientific showed
that the real-time search SPS MS3 method improved identification rates by 30% with higher accuracy and less
interference than the MS2 method by Q Exactive HF-x, Tulane’s existing mass spectrometry system. The
identification rates via the Data-dependent acquisition (DDA) method increased by 76% using Eclipse with
FAIMS Pro interface at the protein level, and increased by 30% using the Data-independent acquisition (DIA)-
based label-free method. The requested electron-transfer dissociation method could better support post-
translation modification analysis with more confident identification. The new generation tribrid MS gives added
experimental throughput while providing depth, coverage, and single-cell proteomics capabilities. These
performance characteristics will be highly useful to the large base of Tulane researchers using advanced LC-
MS/MS analyses for biomedical research.
We have identified an instrument user base of 32 faculty members (12 major users and 20 minor users) funded
by NIH or other federal grants. They have requested the features provided by the proposed instrument to conduct
their studies, demonstrating the significant need for the requested system. Meeting this need, therefore, is
expected to advance multiple high-impact biomedical research projects designed to promote public health across
a wide array of subject areas and NIH research priorities.