Project Summary/Abstract This proposal requests funding for the purchase of a Bruker timsTOF HT mass spectrometer (timsHT MS) coupled with an Evosep One HPLC in order to meet the increasing demand for high-performance proteomics instrumentation at Rutgers University Biomedical and Health Sciences (RBHS)-Newark Campus. The Center for Advanced Proteomics Research (CAPR) at RBHS-New Jersey Medical School in Newark will benefit from this instrument, enhancing service quality. With ever-increasing service needs from NIH-funded researchers, the existing Lumos Orbitrap MS has been exhausted, resulting in a long delay for large-scale proteomics studies. This wait time impedes the progress of NIH-funded investigations. Therefore, adding a timsHT MS at CAPR will speed up the research momentum of Rutgers' NIH-supported researchers. Experienced CAPR scientists will operate the new timsHT MS to conduct wide-scale proteomics studies for a major user group of 15 NIH-funded researchers and others. These investigations aim to analyze more samples quickly and identify more low-abundant proteins, post-translational modifications, and their positional isomers. To achieve these objectives, a high-end MS that can offer sensitivity, speed, and time-saving technology to distinguish isobaric peptides and robustness for boosting instrument uptime. The timsHT MS has a unique blend of engineering and software functionalities that will enable sensitive proteomics studies to meet three unmet needs. Firstly, deep proteome coverage will be achieved through dual Trapped Ion Mobility Spectrometry (TIMS), which facilitates ~30- fold time-focusing of low-level peptide ions, and software that is aware of collision cross-section for in-depth peptide identification and quantification. Secondly, high sensitivity without compromise on speed and resolution will be achieved through Parallel Accumulation - Serial Fragmentation technology, which facilitates MS/MS acquisition at 150 Hz, enabling speedy studies of moderate-level samples with short LC gradients. Thirdly, instrument uptime and robustness will be maximized through an orthogonally positioned ion source to cut impurities in the mass analyzer, a dual-TIMS funnel to filter contaminating gas, and an improved focus on low-level peptide ions. The NIH-funded purchase of this shared timsHT MS will empower CAPR to overcome the time limitations of existing equipment and offer complementary MS technologies to support RBHS-Newark researchers and those from neighboring institutions in carrying out their research goals on NIH-funded grants.