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Expanding experimental scale and increasing sample throughput
Biomarker panels cover a wide range of molecular classes from small molecules, including steroids and drug metabolites, to large molecules, including peptides and proteins.
When developing verified and validated LC-MSn methods, overcome the challenges that each compound class presents with the Thermo Scientific Stellar Mass Spectrometer. With fast polarity switching speeds, hyper-fast tMS2 and tMS3 acquisition speeds using either HCD and CID, Stellar MS extends quantitative performance to effectively and efficiently transition biomarker from discovery to validation.
Translational proteomics: Verify more analytes per unit time
Achieve superior quantitative performance at scale to confidently verify more putative biomarkers with faster LC gradients. The Stellar mass spectrometer combines real-time data acquisition management with hyper-fast acquisition speeds to extend quantitative performance needed to confidently transfer optimal targets to validation.
To ensure reproducible measurements with sensitivity and specificity achieving comprehensive sample coverage, Stellar Mass Spectrometer acquires full scan MS2 data with hyper-fast acquisition speeds, up to 140 Hz, with Thermo Scientific Adaptive RT routine performing dynamic retention time adjustments.
Increase confident peptide verification by 4X over previously published methods to extend biological interpretation for more complex systems. The Stellar mass spectrometer can profile nearly 80% of detectable peptides from extracellular vesicles that map to over 2000 proteins using a 60-minute gradient, expanding the number protein of signaling pathways and the depth of covering in one method.
Stellar MS expands the scale of targeted peptide quantitation for almost 9,000 peptides using a 60-minute gradient. A total of 7,666 peptides were measured with a coefficient of variance < 20%.
Fabregat A, et al. Reactome diagram viewer: data structures and strategies to boost performance. Bioinformatics (Oxford, England). 2018 Apr;34(7) 1208-1214. doi: 10.1093/bioinformatics/btx752. PubMed PMID: 29186351. PubMed Central PMCID: PMC6030826.
Low protein load/single cell proteomics
Experience ultimate instrument flexibility to acquire fast and sensitive untargeted DIA and targeted MS2 (tMS2) acquisition to drive discovery to verification workflow while minimizing sample consumption. To automate method creation, processing, and importing gas-phase fractionated chromatogram libraries to create tMS2 acquisition, Stellar mass spectrometer utilizes Thermo Scientific PRM Conductor suite of tools in Skyline software.
"If I were developing a new targeted biomarker assay, I would use a platform like this as it is a triple quadrupole killer."
Brian Searle
Assistant Professor
The Ohio State University Medical Center
Sample preparation strategy with isolated cell populations with IL-2 and IL-15 stimulated T-cell determined by flow cytometry with percent composition based on gating procedures. Estimated loading amount was less than 1 ng.
Sensitive and rapid targeted immunopeptidomics method development
Advanced instrument performance enables targeted immunopeptidomics to handle the dynamic range, biological matrix, and non-tryptic sequence required to advance method development. Stellar Mass Spectrometer excels at fast and sensitive tMS2 and tMS3 data acquisition using both HCD and CID strategies forming diagnostic product ions to meet experimental requirements.
Quantitative analysis of an endogenous Class I MHC peptide using tMS2.
Quantitative analysis of the same MHC peptide as figure on left using tMS3.
Quantitative analysis of the heavy labeled MHC peptide analogue spiked at 1 amol using tMS2.
Quantitative analysis of the heavy labeled MHC peptide analogue spiked at 1 amol using tMS3.
Related immunopeptidomics resources
Clinical translational research: Democratize targeted methods for routine absolute quantitation for highly multiplexed panels
Expedite routine quantitative method development moving to validation using automated method building and dynamic scheduled retention time control ensuring a sufficient number of data points per LC peak for both the heavy and light peptide analogues. Implementing the Thermo Scientific Adaptive RT routine, Stellar mass spectrometer can maximize the target analyte pairs to maximize sample coverage and quantitative sensitivity while delivering massive throughput.
Enhance pathway mapping confidence with comprehensive site-specific phosphopeptide quantitation using Synchronous Precursor Selection tMS3 acquisition. Intelligent isolation of multiple MS2 product ions, followed by simultaneous dissociation increases quantitative sensitivity, selectivity, and specificity to identify activated pathways. The Stellar mass spectrometer achieves SPS tMS3 acquisition rates up to 40 Hz enabling the full complement of stable isotope-labeled standards and endogenous targets.
Targeted quantitative sensitivity using tMS2 can be limited by background matrix (right side). Performing SPS tMS3 acquisition maximizes sensitivity, suppresses matrix interference, and maintains PTM site-specificity by isolating multiple diagnostic MS2 product ion that generate three key MS3 product ions.
Resources for clinical translational research
Targeted metabolite profiling at untargeted scale
Expand the experimental scale for targeted quantitative metabolite expression profiling using Stellar mass spectrometer. Fast acquisition speeds and parallel ion packet management enables more analytes covering a wider range of compound classes to be quantified while maintaining the sample throughput required for populational studies.
Streamline method development for highly multiplexed targeted metabolite quantitation
Stellar mass spectrometer’s fast polarity switching and tMSn acquisition speed enables comprehensive analyte characterization using LC-MSn charcterization, regardless if reference standards are present. Validated tandem MS spectra can also be imported from the Thermo Scientific mzCloud Mass Spectral Library for confirmation.
Targeted lipidomics: Quantify with confidence
Overcome the limitations of using one SRM/MRM transitions per lipid with Stellar mass spectrometer acquisition methods incorporating tMS2 and tMS3 using both CID and HCD increasing the specificity needed to determine biological reference levels for lipid species.
Go beyond lipid profiling
Triacylglycerides (TAG), critical lipid classes involved in metabolism, are quantified for health assessment. Isomeric TAGs often co-elute using standard chromatographic separation methods requiring MS3 fragmentation to introduce the specificity needed to quantify each isomer.
The Stellar Mass Spectrometer acquires fast tMS2 and tMS3 data up to 140 and 40 Hz, respectively, enabling qualitative and quantitative TAG analysis in one experiment.
New paradigm for targeted small molecule quantitation
Developing routine quantitative methods that maximize sensitivity, selectivity, and specificity for more analytes often requires compromises in productivity. Stellar mass spectrometer uses the QR5+ quadrupole mass filter, dual-purpose ion routing multiple, and dual-pressure linear ion trap mass analyzer to maximize quantitative performance for more compounds in a method that cover a wider range of molecular classes.
The innovative hardware and software advancements deliver fast and sensitive tMS2 and tMS3 acquisition using either beam-type (HCD) or ion trap resonant (CID) dissociation techniques within a single instrument method with drop-down settings to maximize productivity.
Resources for small molecule quantitation
To manage large target compound panels, Stellar MS can reach CID-CID acquisition speeds greater than 30 Hz
To increase quantitative performance for many steroid hormones in the presence of biological matrices, incorporate fast and sensitive resonant CID excitation at both tMS2 and tMS3.
Dihydrotestosterone (DHT) fragmentation is dominated by two neutral-loss product ions based on the hydroxyl and carbonyl functional groups presenting challenges for beam-type fragmentation (HCD), which is similar to the process used on triple quadrupole mass spectrometers. Resonant CID in the high-pressure linear ion trap concentrates the majority of the MS2 product ion signal into the two neutral loss product ions that can be further isolated and fragmented in the linear ion trap generating an MS3 spectrum covering the same product ion mass range as that performed using beam-type fragmentation.
For Research Use Only. Not for use in diagnostic procedures.