Fluorescent Labeling Compounds for Multiplex DNA Assays
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Solution Overview
Problem
Current methods for non-radioactive detection of biological analytes, particularly in multiplex DNA probe assays and DNA sequencing, face challenges with dye label selection due to constraints imposed by electrophoretic separation and enzyme treatment, necessitating a broadly applicable and shelf-stable labeling methodology.
Innovation Solution
Development of compounds with a hydroxy- or protected hydroxy-substituted fluorescent dye linked to an alkanoic acid or heteroalkanoic acid moiety, allowing for versatile labeling of biological agents with emission wavelengths between 400 nm to 1200 nm, including covalent attachment to solid supports and biological agents.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional dye labels are used for multiplex detection, then the detection of multiple analytes is enabled, but the selection of dye labels is constrained by electrophoretic separation and enzyme treatment requirements
Solution Approach 1:
The patent creates a universal labeling platform using carbodiimide chemistry that works with multiple types of fluorescent dyes (carboxyl-, hydroxyl-, and amino-substituted dyes) and biological materials (oligonucleotides, proteins, peptides). This universal approach allows the same labeling methodology to be applied across different dye types and applications, achieving versatility without compromising reliability through standardized reaction conditions and shelf-stable compound design.
2Measurement precision
If radioactive labels are used for detection of biological analytes, then detection sensitivity is achieved, but safety risks and environmental impact increase
Solution Approach 1:
The patent transitions from radioactive to non-radioactive detection by changing the detection parameter from radioactivity to fluorescence. This parameter change maintains detection sensitivity through the use of fluorescent dyes with appropriate emission wavelengths while eliminating safety risks and environmental hazards associated with radioactive materials. The carbodiimide labeling methodology enables this transition by providing efficient covalent attachment of fluorescent dyes to biological analytes.
3Object-affected harmful factors
If fluorescent dyes are used for multiplex detection, then non-radioactive detection is achieved, but the labeling methodology must be broadly applicable to various dyes and biological materials
Solution Approach 1:
The patent introduces carbodiimide chemistry as an intermediary mechanism that facilitates the coupling between carboxyl groups on fluorescent dyes and amino groups on biological materials. This intermediary approach allows the labeling methodology to be broadly applicable to various dye types and biological materials without requiring dye-specific or material-specific protocols, thereby achieving versatility while eliminating radioactivity.
4Stability of the object's composition
If compounds with hydroxy-substituted fluorescent dyes linked to alkanoic acid are developed, then shelf stability and versatility are improved, but compound complexity increases
Solution Approach 1:
The patent creates composite fluorescent labeling compounds by combining hydroxy-substituted fluorescent dyes with alkanoic acid moieties through carbodiimide chemistry. This composite structure integrates the fluorescent properties of the dye with the reactive carboxyl group of the alkanoic acid, achieving both shelf stability and versatility in a single compound design. The composite nature of these compounds allows them to function as stable precursors that can be activated and coupled to various biological materials.
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
Provides a shelf-stable and versatile fluorescent labeling solution compatible with various dyes, enabling efficient labeling of biological agents across different applications, including DNA sequencing and multiplex assays, with improved throughput and reduced uncertainties.
Implementation Method 1
The probes are prepared by covalent attachment of a fluorescent dye to an oligonucleotide
Implementation Method 2
compounds that have an emission wavelength of from about 400 nm to about 1200 nm
Data Source
AI summary
Compounds useful in the fluorescent labeling of biological materials are provided along with methods for their use and preparation.