Direct Mass Technology | Thermo Fisher Scientific

Overview

Unravel molecular complexity faster and drive more confident discoveries

Directly measure the true mass of complex biomolecules with unmatched accuracy—eliminating the ambiguity of traditional ensemble methods. Confidently characterize protein complexes, biotherapeutics, and viral particles that were previously too complex to resolve. The Thermo Scientific Direct Mass Technology mode redefines mass spectrometry by enabling parallel, individual ion charge detection.

Detect subtle variations such as post-translational modifications with exceptional clarity, and gain deeper insight into proteoforms and emerging drug modalities. Powered by a charge-detection enabled Orbitrap mass analyzer, unravel molecular complexity faster and drive more confident discoveries.

Direct Mass Technology mode enables:

Direct mass determination, eliminating need for m/z-based deconvolution
Up to 20-fold increase in resolution relative to traditional ensemble ion measurements
Requires less sample or less concentrated samples.

Direct Mass Technology mode

See how Direct Mass Technology mode adds charge detection for the simultaneous measurement of m/z and z for individual ions

Which mass spectrometer works best for your experiment?

This revolutionary mode is currently available on the Thermo Scientific Q Exactive UHMR Hybrid Quadrupole-Orbitrap mass spectrometers and is in technical preview on the Thermo Scientific Orbitrap Ascend Tribrid Mass Spectrometer.

	Q Exactive UHMR MS	Orbitrap Ascend Tribrid MS
Applications	Best for the largest biological samples, IE assessing empty and full viral capsids.	Assess protein complexes, IE membrane protein mimetics, oligonucleotide complexes, or protein assemblies.
Mass range	Access measurements up to m/z 80,000 and mass measurements in the megadalton range.	Access measurements up to m/z 16,000 and mass measurements in the hundreds of thousand Dalton range.
Fragmentation	Perform MS2 and pseudo MS3 experiments for structural analysis and top-down fragmentation experiments. Use in-source CID for complex degradation and HCD for fragmentation.	Perform MSn (n>2) for structural analysis and top-down fragmentation. Use orthogonal fragmentation techniques ranging from collision-based to radical-based. (including HCD and CID with options for ETD, EThcD, ETciD, and UVPD).

A solution to match the challenge of biological complexity

"One of the most exciting things about Direct Mass Technology mode is it can make the impossible seem very possible. And then, once possible, you get the belief that you can understand protein-level biology. And this is what proteomics needs. We need to understand that we're not overwhelmed by the scale of complexity in the human proteome, that we can actually build tools that match the scale of our biology with the scale of the solution."

Neil Kelleher, Ph.D.
Director, Northwestern Proteomics & the Chemistry of Life Processes Institute
Northwestern University

Direct Mass Technology mode for charge detection

Transform m/z spectrometry to mass spectrometry with simultaneous parallel individual ion measurement of both mass to charge and charge.

Learn more ›

/content/lifetech/north-america/en-us/home/global/forms/industrial/direct-mass-technology-revolutionizing-proteomeform-analysis.html

On-demand Webinar: Unlocking the Power of Direct Mass Technology

Learn how this revolutionary charge detection mass spec technique characterizes and identifies proteoforms using minimal protein quantities.

View webinar ›

Go beyond with native MS

Perform the highest quality native intact mass and top-down analysis in structural biology and biopharma research with the Q Exactive UHMR mass spectrometer.

Learn more ›

Key benefits

Charge determination

The Direct Mass Technology mode adds charge detection capabilities to enable simultaneous measurement of both mass-to-charge (m/z) and charge (z) for hundreds of individual ions in parallel. This allows for the calculation of the mass of the ions directly, without the traditional need for m/z dependent deconvolution that relies on resolved signals in ensemble measurements.

Increase resolution

The Direct Mass Technology mode measures individual ions directly, resulting in 10-20-fold increases in resolution compared to ensemble measurements with the same resolution settings. Individual ion measurements with the Direct Mass Technology mode can allow the isotopic resolution of large and complex analytes typically not possible with ensemble measurements.

Use less sample

Since the Direct Mass Technology mode uses individual ion measurements, the number of ions required for a single measurement is drastically reduced. This means samples can be hundreds of times diluted compared to ensemble measurements, saving precious sample.

Go big

Measurements are read out directly in the mass domain with the Direct Mass Technology mode, enabling measurements up to 80,000 m/z and mass measurements in the megadalton range. Go beyond the capabilities of native ensemble measurements to explore the redefined limits of addressable size and complexity.

Overview of the Direct Mass Technology mode workflow

Available on the Q Exactive UHMR mass spectrometer, the Direct Mass Technology mode adds:

Automated ion population control to maximize throughput and signal quality for parallel individual ion measurements
A charge calibration procedure
STORIboard processing software to perform charge calibration, data processing, and data visualization developed in collaboration with Proteinaceous, Inc.

Click image to enlarge

Step 1: As with traditional Orbitrap measurements, the mass to charge ratio m/z is determined by the frequency of axial rotation around the central electrode.

Click image to enlarge

Step 2: With the Direct Mass Technology mode every ion is also analyzed individually in parallel. The integrated signal for each individual ion is plotted over time through measuring the rate of the induced charge on the outer electrode at the specific frequency of the ion and is known as the Selective Temporal Overview of Resonant Ions (STORI).

Click image to enlarge

Step 3: The slope of the STORI plot for each individual ion is then compared to the charge calibration curve to determine the charge (z). The mass of each individual ion can then be directly calculated using the mass to charge (m/z) and charge (z) information generated by the measurements.

The Direct Mass Technology mode adds extra dimensionality to Orbitrap measurements with parallel individual ion measurements to produce high-resolution results directly in the mass domain.

Applications

Glycoform analysis

Click image to enlarge

Reveal exquisite details

Standard native ensemble measurements can be restricted by spatial resolution and dynamic range, resulting in limited detection of glycoforms. With the Direct Mass Technology mode more masses can be resolved through simultaneous charge detection to more than double the number of uniquely assigned glycoforms.

Membrane proteins and complexes

Click image to enlarge

Increase sensitivity and resolution

Membrane proteins and complexes require considerable sample preparation and have unique analytical challenges. The Direct Mass Technology mode provides higher sensitivity and resolution through individual ion measurements, allowing for less sample consumption and delivering confidence in results from a single analysis.

Biotherapeutic characterization

Click image to enlarge

Turn complexity into clarity

Complex biotherapeutics present analytical challenges that can require multiple experiments and very careful sample processing and handling. The Direct Mass Technology mode allows for charge state and mass determination to reveal a wider molecular weight distribution and more detailed insights into complex analytes.