Mass Spectrometry Data Acquisition with Dynamic Isolation Windows

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Solution Overview

Problem

Current mass spectrometry techniques, such as data-dependent and data-independent acquisition methods, fail to provide comprehensive lists of all proteins or peptides in complex biological samples, particularly missing low-abundance peptides due to limitations in chromatographic separation and fragmentation analysis.

Innovation Solution

A hybrid method combining aspects of data-dependent and data-independent mass spectrometry, involving measurement cycles with survey mass spectra and data-independent product-ion analyses, where precursor ions are isolated within restricted windows, fragmented, and analyzed, with adjustable parameters to maximize spectral quality and correlate parent and fragment ions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If data-dependent acquisition is used to select and isolate single ion species for fragmentation, then identification accuracy is improved, but low-abundance peptides are missed due to focus on abundant ions

Engineering Contradiction:
Improveidentification accuracyVSAvoidpeptide coverage
Core Design Contradiction:
Measurement precisionVSQuantity of substance

Solution Approach 1:

The mass spectrum is segmented into multiple isolation windows covering different m/z ranges. Each window is processed independently to identify precursor ions, ensuring both abundant and low-abundance peptides are captured across the full spectral range rather than focusing on a single dominant ion population.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The method dynamically adjusts survey scan parameters based on real-time spectral attributes. When low-abundance ions are detected in certain m/z regions, the system increases survey scan frequency or adjusts isolation window parameters to enhance detection sensitivity for these lesser-abundance species while maintaining overall identification accuracy.

Inventive Principle:
Principle #15Dynamics

2Ease of operation

If survey mass spectra are obtained at regular time intervals, then data acquisition is simplified, but spectral quality and peptide detection are compromised due to fixed parameters

Engineering Contradiction:
Improvedata acquisition simplicityVSAvoidspectral quality
Core Design Contradiction:
Ease of operationVSMeasurement precision

Solution Approach 1:

The system continuously monitors spectral attributes from survey scans and uses this feedback to dynamically adjust acquisition parameters. When spectral complexity increases or specific m/z regions show high ion density, the system automatically modifies survey scan frequency, isolation window width, or fragmentation parameters to optimize spectral quality without manual intervention.

Inventive Principle:
Principle #23Feedback

Solution Approach 2:

Acquisition parameters such as survey scan frequency, isolation window width, and fragmentation collision energy are changed based on real-time spectral characteristics. The system transitions between different parameter sets depending on detected spectral attributes, enabling adaptive optimization of both simplicity and quality.

Inventive Principle:
Principle #35Parameter changes

3Stability of the object's composition

If fixed isolation windows are used for product-ion analysis, then analysis consistency is maintained, but interference from co-isolated ions increases

Engineering Contradiction:
Improveanalysis consistencyVSAvoidion interference
Core Design Contradiction:
Stability of the object's compositionVSObject-affected harmful factors

Solution Approach 1:

Isolation window parameters are dynamically adjusted based on the detected distribution and abundance of precursor ions in each m/z region. When ions are densely packed, the system narrows isolation windows or increases spacing to reduce co-isolation of interfering ions, while maintaining consistent analytical coverage across the full m/z range.

Inventive Principle:
Principle #15Dynamics

Solution Approach 2:

Different isolation window parameters are applied to different m/z regions based on local ion density and spectral characteristics. High-density regions receive narrower or more spaced windows to minimize interference, while low-density regions use standard parameters, ensuring optimal performance across heterogeneous spectral landscapes.

Inventive Principle:
Principle #3Local quality

4Quantity of substance

If comprehensive survey scans covering full m/z range are performed, then all peptide types are detected, but analysis time increases significantly

Engineering Contradiction:
Improvepeptide coverageVSAvoidanalysis time
Core Design Contradiction:
Quantity of substanceVSLoss of time

Solution Approach 1:

The full m/z range is divided into multiple isolation windows that are processed in parallel or sequential batches. This segmentation enables comprehensive coverage of all peptide types while reducing the time required for each individual survey scan and product-ion analysis cycle, as the system can efficiently cycle through windows without requiring full-range scans at maximum resolution.

Inventive Principle:
Principle #1Segmentation

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This approach enhances the detection of a broader range of peptides by optimizing survey and product-ion analysis parameters, improving the identification and quantification of peptides, including low-abundance species, and reducing interference, thereby providing more comprehensive proteome coverage.

Implementation Method 1

isolation of precursor ions within a restricted isolation window

Methodology Applied
Scientific EffectElectromagnetic field separation: Electric Field

Implementation Method 2

subjecting such so-isolated precursor ions to fragmentation so as to yield product ions... collision-induced dissociation (CID)... kinetic energy is imparted to the peptide ions, whereby the introduced energy is converted into internal vibrational energy

Methodology Applied
Scientific EffectCollision-induced dissociation: Impact Force

Implementation Method 3

mass analysis of the fragment ions generated from the precursor ions that were isolated in the corresponding isolation window

Methodology Applied
Scientific EffectIon cyclotron resonance: Magnetic Field

Data Source

PatentEP3340275B1Data-independent mass spectral data acquisition including data-dependent precursor-ion surveys
Publication Date: 2021.11.03 THERMO FINNIGAN LLC
  • EP3340275B1 patent drawingFigure 1A
  • EP3340275B1 patent drawingFigure 1B
  • EP3340275B1 patent drawingFigure 1C

AI summary

A mass spectrometry method comprises: acquiring a series of survey mass spectra of first-generation ions generated from a sample; acquiring a series of fragment-ion mass spectra, each being a record of a respective set of fragment-ion species generated by fragmentation of a respective subset of the first-generation ions within a respective mass-to-charge isolation range; adjusting mass spectrometer operational parameters used to acquire a later one of the survey mass spectra based on results of an earlier one of the survey mass spectra; dividing the acquired series of fragment-ion mass spectra into a first group wherein an appearance of a fragment-ion species correlates with the appearance of a first-generation ion species observed in a survey mass spectrum and a second group wherein no obvious correlation is observed between fragment-ion species and first-generation ion species; and mathematically processing the spectra of the first and second groups by different mathematical procedures.