Orbitrap Astral to support investigator proteomics research in a Core facility.

Yale University, New Haven CT

Investigators

Abstract

7. Project Summary/Abstract Funding is requested for the purchase of the current state-of-the-art Orbitrap Astral mass spectrometer system. The Orbitrap Astral is the newest hybrid (quadrupole/Orbitrap/Astral) MS platform in its class with a novel Astral mass analyzer capable of fragment ion scanning above 200 Hz at a resolution of 80,000 at m/z 524, a leading-edge Orbitrap mass analyzer which enables scan speeds of up to 40 Hz at resolution up to 480,000 at 200 m/z, and sub-part-per-million mass accuracy. The ability to parallelize precursor/fragment ion detection in the two mass analyzers along with the ultrafast scan rates offered by this cutting-edge instrument enable >5-fold more MS/MS scans/second as compared to the best mass spectrometers currently housed in the Keck MS & Proteomics Resource, enabling incredibly deep coverage from single-shot proteomics experiments. The Orbitrap Astral MS will be coupled to a Vanquish Neo UHPLC system that has excellent peak separation efficiency and reproducibility, and the proposed MS system will have all the necessary hardware and software needed for rapid identification, profiling, and quantitative proteomics workflows. Based on the manufacturer’s data, we expect this instrument will be able to identify over 9,000 proteins in a 15-minute LC gradient or 12,000 proteins in a 60-minute gradient for complex mammalian extracts. This unprecedented depth of coverage over short gradients would enormously improve the Resource’s sample throughput for biomarker discovery, helping to minimize the current 3+ week sample backlog and ensuring that the Resource has the state-of-the-art mass spectrometry capabilities to accommodate additional investigators who would benefit from these advanced technologies. Addition of the Orbitrap Astral will also substantially increase sensitivity for the identification of low-level proteins and their post translational modifications and allow the Resource to push the envelope of single-cell proteomics. The advances incorporated into this instrument make it the ideal platform to support the LFQ, DIA, and PTM workflows that have become highly-requested services by the 142+ Yale and non-Yale investigators submitting >7,668 samples to the MS & Proteomics Resource in FY23 and FY24 to date. This application is supported by 28 investigators encompassing 8+ different disciplinary research fields and 40+ different projects, and the proposed eight Major users of the Orbitrap Astral MS include members of four NIH and National Centers at Yale (the Yale/NIDA Neuroproteomics Center, the Yale Cancer Center, the Yale Alzheimer’s Disease Research Center, and the Interstitial Lung Disease Center of Excellence). The strengths of this proposal are manifold and include continued very strong institutional support not only in the initial committed investment (with >$227K committed towards service maintenance costs), but also in long-term assurances to cover any deficit that may occur for the Keck MS & Proteomics Resource; the sustained, balanced operational budget of the Resource; and the continued usage of and productive collaboration with the Resource by hundreds of NIH-funded investigators annually.