Introduction
Speed and sensitivity in combination with high-resolution and accurate mass (HRAM) remain key to further propel large cohort studies and advance single-cell proteomics to the next level. This need is addressed in this work by a novel instrument platform that combines the Thermo Scientific™ Orbitrap™ analyzer with the novel Thermo Scientific™ Astral™ analyzer. The combination of these two analyzers allows for unprecedented performance that raises mass spectrometry to the next level.
Methods
The Thermo Scientific™ Orbitrap™ Astral™ mass spectrometer allows for acquisition of HRAM MS1 spectra with up to 480K resolution in the Orbitrap analyzer and, in parallel, HRAM MS2 spectra at up to 200Hz in Astral analyzer. In total, 5 ion packages are processed in parallel to maximize the usage of ions and allow for unprecedented sensitivity and speed so that an entire MS/MS cycle can be executed in under 5ms.
The new platform and its key components that enable speed, sensitivity, mass accuracy and resolving power of up to 100K in the Astral analyzer will be presented. Special emphasis will be given to the ion handling, ion processor, Astral analyzer, and high-dynamic range detector which allows for single-ion detection efficiency.
Results
For performance characterization, the new platform has been characterized with Pierce™ Flexmix™ calibration solution, HeLa digest and proteins. For 200ng of HeLa digest, this resulted in >7,000 protein groups identified at 300 samples-per-day (SPD), >8000 at 180 SPD, and >10,000 at 30 SPD. For low sample load (250pg HeLa), >6000 protein groups can be detected when processed in Spectronaut 18.
Conclusion
A novel instrument with unique new technologies represents a significant improvement that will help to bring proteomics to the next level