Isobaric Mass Labels for Multi-Analyte Quantitation

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Solution Overview

Problem

Current methods for biomarker validation and quantitation in mass spectrometry are limited by the need for high specificity reagents like antibodies, which are costly, time-consuming to produce, and can only detect a limited number of analytes simultaneously, especially when dealing with proteins across wide concentration ranges.

Innovation Solution

A method using isobarically tagged reference biomolecules that allow for multi-point standard curves in mass spectrometry, enabling the simultaneous quantitation of multiple analytes by differentially labeling test and calibration samples with isobaric mass labels, allowing for calibration and accurate quantitation without increasing MS complexity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If antibody-based methods are used for biomarker validation, then specificity and accuracy are improved, but cost and time consumption increase significantly

Engineering Contradiction:

ImprovespecificityVSAvoidtime consumption

Core Design Contradiction:

Measurement precisionVSLoss of time

Solution Approach 1:

The patent replaces expensive, time-consuming antibody reagents with inexpensive, rapidly synthesized small molecule probes that can be produced in high throughput. These small molecule probes serve as disposable alternatives to antibodies, eliminating the need for costly purification and validation processes while maintaining detection capability through mass spectrometry.

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent substitutes the biological recognition mechanism (antibody-antigen binding) with a chemical recognition system based on small molecule-probe interactions detected by mass spectrometry. This replacement transitions from a complex biological system requiring extensive validation to a streamlined chemical analysis system with rapid turnaround.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

2Measurement precision

If antibody-based methods are used for biomarker detection, then detection accuracy is improved, but the number of analytes that can be detected simultaneously is limited

Engineering Contradiction:

Improvedetection accuracyVSAvoidnumber of analytes detected simultaneously

Core Design Contradiction:

Measurement precisionVSProductivity

Solution Approach 1:

The patent employs a universal mass spectrometry-based detection platform that can analyze multiple different analytes using the same instrumental system. By using small molecule probes with distinct mass signatures, the system achieves multi-functionality, allowing simultaneous detection of numerous biomarkers in a single experiment without requiring separate antibody validations for each analyte.

Inventive Principle:

Principle #6Universality (Multi-functionality)

Solution Approach 2:

The patent utilizes mass-to-charge ratio as a distinguishing parameter for identifying different analytes. By assigning unique mass signatures to different small molecule probes through isotopic labeling or structural variation, the system can differentiate and quantify multiple analytes simultaneously based on their distinct mass parameters, overcoming the limitation of antibody-based methods.

Inventive Principle:

Principle #35Parameter changes

3Ease of operation

If traditional mass spectrometry methods are used for protein quantitation, then simplicity of method is maintained, but sensitivity and dynamic range are insufficient

Engineering Contradiction:

ImprovesimplicityVSAvoidsensitivity

Core Design Contradiction:

Ease of operationVSMeasurement precision

Solution Approach 1:

The patent introduces small molecule probes as intermediary agents that bind to target proteins and provide enhanced mass spectrometric signals. These probes act as mediators that amplify the detectability of proteins, improving sensitivity without complicating the overall workflow. The probes are detected alongside their protein targets through the same mass spectrometry system, maintaining operational simplicity.

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

Enables the measurement of up to 10 to 50 analytes in a single LC-MS/MS experiment with improved accuracy and sensitivity, overcoming the limitations of antibody-based methods and expanding the dynamic range of protein concentrations.

Implementation Method 1

determining by mass spectrometry the quantity of the analyte in the test sample and the quantity of analyte in each aliquot in the calibration sample

Methodology Applied

Scientific EffectMass spectrometry:

Implementation Method 2

measurement of up to 10 to 50 analytes in a single LC-MS/MS experiment

Methodology Applied

Scientific EffectLiquid chromatography: Chromatography

Data Source

PatentUS10976321B2Mass spectrometric quantitation

Publication Date: 2021.04.13 ELECTROPHORETICS LTD

AI summary

Provided is a method of assaying for an analyte, including combining a test sample having the analyte, with a calibration sample having at least two different aliquots of the analyte, each aliquot having a different known quantity of the analyte. The test sample and each aliquot are differentially labeled with one or more isobaric mass labels each with a mass spectrometrically distinct mass marker group, such that the test sample and each aliquot of the calibration sample can be distinguished by mass spectrometry. The method further includes determining by mass spectrometry the quantity of analyte in the test sample and in each aliquot, and calibrating the quantity of analyte in the test sample against known and determined quantities of analytes in the aliquots.