Fluorescence Quencher Polymer Conjugates for Phycoerythrin
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Solution Overview
Problem
Current fluorescence quenching compounds are ineffective in interacting with and quenching the strong fluorescent emissions from macromolecular emitters like phycoerythrin, which is a red protein-pigment complex present in red algae and cryptophytes.
Innovation Solution
Development of a fluorescence quenching compound comprising discrete fluorescence quenching moieties such as EQ0, EQ1, and EQ2 chemically conjugated to a non-protein polymer like branched polyethylenimine, which enhances quenching efficiency by forming stable amide bonds with primary amines, particularly useful for macromolecular emitters like phycoerythrin.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If standalone quenching moieties are used, then the device complexity is low, but the fluorescence quenching efficiency is insufficient for strong macromolecular emitters
Solution Approach 1:
The patent applies composite materials by combining multiple quenching moieties (such as EQ0, EQ1, EQ2) with a polymer backbone (branched polyethylenimine) to create a hybrid quencher molecule. This composite structure leverages the high quenching efficiency of multiple chromophores while the polymer framework provides stability and multiple interaction sites, resolving the contradiction between simplicity and effectiveness.
Solution Approach 2:
The patent merges multiple discrete quenching moieties into a single polymeric quencher molecule through chemical conjugation. By combining several quenching units (EQ0, EQ1, EQ2) with the polyethylenimine backbone, the system achieves cumulative quenching effects that overcome the limitations of single-moiety quenchers against strong macromolecular fluorophores like phycoerythrin.
2Reliability
If multiple quenching moieties are conjugated to polymer, then the fluorescence quenching efficiency increases, but the manufacturing process becomes more complex
Solution Approach 1:
The patent employs preliminary action by pre-functionalizing the polyethylenimine backbone with reactive groups (such as NHS esters or isocyanates) before conjugation. This allows for controlled, stepwise attachment of quenching moieties under mild conditions, simplifying the overall manufacturing process despite the multi-component nature of the final product.
Solution Approach 2:
The patent utilizes parameter changes by adjusting reaction conditions (pH, temperature, solvent composition) to optimize the conjugation process. By controlling these parameters, the patent achieves efficient coupling of multiple quenching moieties to the polymer backbone while maintaining product stability and minimizing side reactions, thereby easing manufacturing complexity.
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
The conjugated quenching compound significantly reduces the fluorescent emission of phycoerythrin to less than 2% of its original level, demonstrating enhanced quenching efficiency compared to standalone quenching moieties, with specific interactions influenced by salt bridges and polymer charge density.
Implementation Method 1
chemically conjugated to a discrete site of the polymer
Implementation Method 2
quenching the fluorescent emissions from strong macromolecular emitters of fluorescence such as phycoerythrin
Implementation Method 3
specific interactions influenced by salt bridges and polymer charge density
Data Source
AI summary
The invention provides chemical conjugates comprising a plurality of fluorescence quenching moieties conjugated to polymer and the use of such conjugates to quench the emission of fluorescence from emitters including macromolecular fluorescent emitters such as phycoerythrin.