Targeted Top-Down Mass Spectrometry for Precursor-Fragment Correlation

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Solution Overview

Problem

Current chromatography and mass spectrometry systems for targeted top-down discovery face limitations in effectively analyzing complex biological molecules, particularly in correlating precursor and product ions and determining the structure of complex molecules due to variations in mass resolution and retention times.

Innovation Solution

The system employs a dual mass analyzer setup with data processing to match and average chromatographic and mass spectrometric properties across multiple data sets, generating inclusion lists and fragmentation parameters to selectively fragment and analyze biopolymers based on mass-to-charge ratios and retention times, enabling detailed analysis of biopolymers like proteins, peptides, and lipids.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If multiple mass analyzers with different resolutions are used to analyze biopolymers, then the measurement precision and completeness of molecular analysis is improved, but the device complexity and data processing difficulty increase

Engineering Contradiction:
Improvemass resolutionVSAvoiddual mass analyzer setup
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The system divides the mass analysis task into segments by using multiple mass analyzers (e.g., quadrupole, time-of-flight, orbitrap) operating at different mass resolutions. Each analyzer handles specific mass ranges or resolution requirements, allowing comprehensive analysis of biopolymers without requiring a single overly complex instrument to perform all functions simultaneously.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The mass spectrometry system is designed with multi-functional capability by integrating multiple types of mass analyzers that can each perform different analytical functions. The system can switch between or combine high-resolution and low-resolution analysis modes, enabling a single platform to address diverse analytical needs for structure elucidation, quantification, and identification of biological molecules.

Inventive Principle:
Principle #6Universality (Multi-functionality)

2Measurement precision

If data from multiple mass chromatographic data sets at different resolutions are averaged, then the accuracy of chromatographic and mass spectrographic properties is improved, but the data processing time and computational complexity increase

Engineering Contradiction:
Improveaccuracy of chromatographic and mass spectrographic propertiesVSAvoiddata processing time
Core Design Contradiction:
Measurement precisionVSLoss of time

Solution Approach 1:

The system performs preliminary processing of mass chromatographic data sets by organizing and pre-aligning data from multiple analyzers before averaging. This includes preliminary matching of retention times and mass-to-charge ratios, and preprocessing to identify corresponding peaks across different resolutions, reducing the computational burden during the final averaging step.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The data processing approach involves changing parameters such as retention time alignment tolerances, mass-to-charge ratio matching thresholds, and weighting factors applied to different data sets during averaging. By optimizing these parameters, the system achieves accurate property determination while managing computational complexity and processing time effectively.

Inventive Principle:
Principle #35Parameter changes

3Measurement precision

If targeted top down discovery is used to analyze complex biopolymers, then the structural determination accuracy is improved, but the difficulty of detecting and measuring precursor and product ions increases

Engineering Contradiction:
Improvestructural determination accuracyVSAvoidcorrelation of precursor and product ions
Core Design Contradiction:
Measurement precisionVSDifficulty of detecting and measuring

Solution Approach 1:

The system employs feedback mechanisms in the data processing workflow where information from product ion fragmentation patterns is fed back to refine the identification and characterization of precursor ions. This iterative process allows the system to use fragmentation data to confirm precursor identities and resolve ambiguities in detecting and correlating ion relationships, thereby improving structural determination accuracy.

Inventive Principle:
Principle #23Feedback

Solution Approach 2:

The system uses intermediate data structures and processing steps as mediators between raw mass spectrometry data and final structural conclusions. These intermediaries include processed chromatographic data, aligned mass spectra, and intermediate fragmentation patterns that facilitate the correlation of precursor and product ions by providing structured, organized information that reduces the complexity of direct ion correlation analysis.

Inventive Principle:
Principle #24Intermediary (Mediator)

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 resolution and accuracy of molecular analysis by harmonizing data from different mass analyzers, allowing for precise identification and sequencing of biopolymers, improving the structural determination of complex molecules.

Implementation Method 1

The eluent can be directed to a mass spectrometer, where the molecules can be detected and a mass can be determined

Methodology Applied
Scientific EffectMass spectrometry:

Implementation Method 2

a sample can be separated based on physical properties of the constituent molecules, such as hydrophobicity, charge, isoelectric point, and the like using liquid chromatography

Methodology Applied
Scientific EffectChromatography: Chromatography

Implementation Method 3

further information can be obtained for particular molecules by fragmenting the molecule and determining the mass of the individual fragments in what is known as an MS/MS experiment

Methodology Applied
Scientific EffectIon fragmentation:

Data Source

PatentEP3136094B1System and method for targeted top down discovery
Publication Date: 2023.11.29 THERMO FINNIGAN LLC
  • EP3136094B1 patent drawingFigure 1
  • EP3136094B1 patent drawingFigure 2
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AI summary

A system for analyzing a sample includes a chromatographic device, a mass resolving device, and a data processor. The chromatographic device is configured to separate components of the sample using a chromatographic column. The mass resolving device is configured to characterize mass spectrographic properties of the separated components in an intact state, and fragment the separated components and characterize mass spectrographic properties of the resulting fragments. The data processor is configured to average chromatographic and mass spectrographic properties from a plurality of mass chromatographic data sets, generate an inclusion list identifying components for fragmentation, instruct the chromatographic device to repeat the separation of the sample and instruct the mass resolving device to fragment the components and characterize the mass spectrographic properties of the fragments, and identify at least one component based on the mass spectrographic properties of the intact state and the corresponding fragments.