Modified Monosaccharide Bacterial Labeling Method
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Solution Overview
Problem
Existing methods for labeling living bacteria, particularly Gram-negative bacteria, are limited in their ability to incorporate non-endogenous monosaccharide compounds into the bacterial envelope, restricting their applicability to a broader range of bacteria, including Gram-positive bacteria.
Innovation Solution
A method involving the use of modified monosaccharide compounds with a first reactive chemical group, such as azido or alkyne, which can be incorporated into the polysaccharides of both Gram-negative and Gram-positive bacteria, allowing for specific labeling through a chemical reaction with a labeling molecule, enabling covalent linkage and detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If modified endogenous monosaccharide compounds (ulosonic acid type) are used for labeling, then Gram-negative bacteria can be specifically labeled, but Gram-positive bacteria cannot be labeled
Solution Approach 1:
The patent develops a universal labeling method using modified non-endogenous monosaccharide compounds that can label both Gram-negative and Gram-positive bacteria, eliminating the need for different labeling strategies for different bacterial types. The modified monosaccharide compounds with reactive groups (azide, alkyne, or vinyl) can be incorporated into the envelopes of diverse bacteria through their biosynthetic pathways, achieving broad applicability across bacterial species.
2Productivity
If endogenous monosaccharide analogs are used, then incorporation into bacterial envelope is efficient, but the method is limited to specific bacterial categories
Solution Approach 1:
The patent changes the chemical parameters of the monosaccharide compounds by introducing reactive groups (azide, alkyne, or vinyl) at specific positions (C-3, C-4, or C-6) of the sugar ring. These parameter changes allow the compounds to maintain their ability to be incorporated into bacterial envelopes through natural biosynthetic pathways while enabling subsequent chemical labeling reactions, thus achieving both efficiency and versatility.
3Adaptability or versatility
If non-endogenous monosaccharide compounds are used, then broader bacterial coverage is achieved, but incorporation into envelope is difficult
Solution Approach 1:
The patent uses modified non-endogenous monosaccharide compounds as intermediaries that can be incorporated into bacterial envelopes through the bacteria's natural biosynthetic pathways. These compounds serve as mediators between the external labeling agents and the internal bacterial structures, allowing efficient incorporation without requiring complex genetic modification of the bacteria. The reactive groups on these intermediary compounds then enable subsequent chemical labeling.
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 method allows for rapid and specific labeling of a wide range of bacteria, including both Gram-negative and Gram-positive species, facilitating fast detection and isolation, overcoming the limitations of previous techniques which were primarily suited for Gram-negative bacteria.
Implementation Method 1
capable to react with a second reactive group of a labeling molecule... generating the reaction of said first reactive group with said second reactive group of said labeling molecule, resulting in a covalent link
Data Source
AI summary
The invention relates to a method for labeling specifically living bacteria, comprising the steps of: a) incubating said bacteria of said sample with at least one modified monosaccharide compound comprising a first reactive chemical group capable to chemically react with a second reactive group, so that a residue bearing said first reactive group is incorporated into the envelope of such bacteria, and b) contacting said modified monosaccharide residue incorporated within the envelope of the bacteria with a labeling molecule comprising a said second reactive group, for generating the chemical reaction of said first reactive group with said second reactive group, characterized in that said modified monosaccharide compound has the following formula (I), or a salt thereof: Wherein —A, B and C can be independently H, OH, NH2, OH and NH2 being substituted or not by protecting groups thereof and —D is an alkyl chain in C2 to C4, each carbon being substituted or not by OH or NH2, OH and NH2 being substituted or not by protecting groups thereof and —at least one of A, B, C or D groups is substituted by a said first reactive group.