Sugar Chain Fluorescent Labeling via Aromatic Amine Concentration
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Solution Overview
Problem
Current methods for fluorescent labeling of sugar chains are inefficient, leading to incomplete labeling and reduced sensitivity in analysis, particularly when using mass spectrometry, due to low labeling efficiency and complexity in peak analysis.
Innovation Solution
A method involving the use of a sugar chain fluorescent labeling technique that reacts sugar chains with a fluorescent substance, such as aromatic amines like 2-aminobenzamide, at a concentration of 0.5 mol/L or more, utilizing a sugar chain capturing carrier to enhance labeling efficiency and specificity, and optimizing conditions for reductive amination reactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional fluorescent labeling methods are used with lower concentrations of aromatic amine, then the reaction conditions are milder and easier to control, but the labeling efficiency is insufficient leading to incomplete labeling and reduced analysis sensitivity
Solution Approach 1:
The patent applies parameter changes by increasing the concentration of aromatic amine to 0.5 mol/L or more, which fundamentally alters the reaction kinetics and thermodynamics to achieve complete labeling. This parameter adjustment resolves the contradiction by ensuring all sugar chains are labeled uniformly, eliminating peak complexity in mass spectrometry while maintaining analytical control through optimized reaction conditions
2Manufacturing precision
If high concentration of fluorescent substance is used to improve labeling efficiency, then labeling completeness increases, but the cost of reagents and complexity of procedure increase
Solution Approach 1:
The patent employs preliminary action by using a sugar chain capturing carrier to isolate and concentrate sugar chains before the fluorescent labeling reaction. This pre-concentration step allows the subsequent high-concentration aromatic amine reaction to be more efficient and controlled, reducing procedure complexity while achieving complete labeling
Solution Approach 2:
The sugar chain capturing carrier acts as an intermediary that facilitates the labeling process. It binds sugar chains selectively, enables concentration, and provides a controlled environment for the high-concentration aromatic amine reaction, thereby simplifying the overall procedure while ensuring complete labeling
3Measurement precision
If reductive amination labeling is performed to achieve high sensitivity in HPLC and CE analysis, then detection sensitivity improves, but the time and labor required for purification and labeling procedures increase significantly
Solution Approach 1:
The patent merges the purification and labeling steps into a single integrated process using the sugar chain capturing carrier. Sugar chains are captured, concentrated, and labeled in sequence without intermediate purification steps, thereby maintaining high detection sensitivity through complete labeling while dramatically increasing sample preparation throughput by eliminating time-consuming separate procedures
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 allows for efficient and easy fluorescent labeling of sugar chains, improving analysis sensitivity and reducing peak complexity, enabling more accurate and rapid analysis by HPLC or mass spectrometry.
Implementation Method 1
it is necessary to label the reducing terminal of a sugar chain by reductive amination and the like in order to obtain high sensitivity
Implementation Method 2
a sugar chain capturing carrier that captures sugar chain molecules
Implementation Method 3
labeling a sugar chain by reacting a sugar chain with a fluorescent substance including an aromatic amine
Data Source
AI summary
An object of the present invention is to provide a simple method for fluorescent labeling of sugar chains, and a sugar chain capturing carrier used therein. According to the present invention, a sugar chain fluorescent labeling method is provided in which sugar chains are captured, recovered and purified by primary amino groups in a sugar chain capturing carrier having the primary amino groups that is used to capture sugar chains, and the captured sugar chains are released from the carrier and then labeled with a fluorescent substance including aromatic amine at a concentration of 0.5 mol/L or more.