Activated Procaine Labeling for Stable N-Glycan Analysis
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Solution Overview
Problem
There is a need for rapid and sensitive labeling methods for N-glycans and amine-containing compounds that do not cause degradation under mild conditions, suitable for high sensitivity detection by fluorescence and mass spectrometry.
Innovation Solution
The use of Activated Procaine or its solvates and salts for labeling N-glycans, amine-containing amino acids, peptides, and proteins, which provide superior detection by fluorescence and equivalent sensitivity by mass spectrometry, allowing rapid tagging and analysis through chromatography and mass spectrometry.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional labeling methods are used for N-glycans, then labeling can be achieved, but the labeling process is slow and requires harsh conditions that cause degradation of labile components
Solution Approach 1:
The patent changes the chemical parameters of the labeling reagent by using activated esters (NHS esters, isothiocyanates, sulfonyl fluorides) instead of conventional reagents. This parameter change enables the labeling reaction to proceed rapidly under mild physiological conditions (pH 7-9, room temperature or 37°C), thereby achieving high productivity while preventing degradation of labile N-glycan structures
Solution Approach 2:
The patent introduces activated ester intermediaries (compounds of Formula I containing leaving groups such as NHS esters, isothiocyanates, or sulfonyl fluorides) that mediate the labeling reaction. These intermediaries react rapidly with the reducing end amino group of N-glycans under mild conditions, enabling fast labeling without harsh conditions that would degrade labile components
2Measurement precision
If conventional labeling reagents are used, then labeling can be performed, but the sensitivity of detection by fluorescence and mass spectrometry is insufficient
Solution Approach 1:
The patent changes the chemical structure parameters of the labeling reagent by incorporating fluorogenic groups and mass spectrometry-enhancing moieties into the activated ester framework. This enables simultaneous high sensitivity detection by both fluorescence and mass spectrometry, achieving superior measurement precision without significantly increasing method complexity
Solution Approach 2:
The patent designs compounds of Formula I that are multi-functional: they serve as both the labeling reagent (through the activated ester group) and the detection tag (through fluorogenic and mass spectrometry-active groups). This universal design enables a single reagent to provide both labeling and high-sensitivity detection functions, eliminating the need for separate detection steps
3Speed
If rapid labeling is achieved using activated esters, then labeling speed improves, but the stability of the labeling reaction may be compromised
Solution Approach 1:
The patent optimizes the parameters of the activated ester leaving groups (NHS esters, isothiocyanates, sulfonyl fluorides) to achieve the right balance: these groups are sufficiently reactive to enable rapid labeling kinetics but form stable amide, thiourea, or sulfonamide bonds with the N-glycan amino group, ensuring labeling stability. The reactions are conducted at controlled pH (7-9) and temperature to maintain both speed and reliability
Solution Approach 2:
The patent uses activated ester intermediaries that are designed to be consumed in the labeling reaction, forming stable final products. The activated ester groups (NHS esters, isothiocyanates, sulfonyl fluorides) are sufficiently reactive to drive rapid labeling but are consumed in the process, forming stable bonds that ensure long-term reliability of the labeled N-glycans
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
Activates Procaine enables efficient and stable labeling of amine-containing compounds, providing enhanced detection sensitivity and rapid analysis, especially for N-glycans, with improved fluorescent and mass spectrometry results compared to existing labels.
Implementation Method 1
reacting said N-glycan, amine-containing amino acid, peptide, protein, or other amine-containing compound with a compound ('Activated Procaine') thereby labeling said N-glycan, amine-containing amino acid, peptide, protein, or other amine-containing compound in the sample
Implementation Method 2
allowing rapid tagging and analysis through chromatography and mass spectrometry
Implementation Method 3
provide superior detection by fluorescence and equivalent sensitivity by mass spectrometry
Data Source
AI summary
The invention relates to an activated form of procaine, and the use of the activated procaine, or salts or solvates thereof, to label amine-containing compounds, such as N-glycans, amine-containing amino acids, amine-containing peptides, amine-containing proteins, or other amine-containing compounds in a sample. Use of activated procaine as a label allows for sensitive detection of compounds labeled with it by both fluorescence and mass spectrometry.