Fluoreno[4,5-cde]oxepine Polymers for Analyte Detection

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Solution Overview

Problem

Existing fluorescent polymers face challenges such as autofluorescence interference, reduced quantum efficiency upon conjugation, self-quenching, and instability, limiting their effectiveness in biological applications, particularly in water-soluble forms.

Innovation Solution

The development of fluoreno[4,5-cde]oxepine-based polymers, which exhibit high fluorescence quantum yield, red-shifted emission, high water solubility, linearity, planarity, and photostability, along with efficient fluorescence resonance energy transfer, by crosslinking the benzene rings, addressing the limitations of unsubstituted polyfluorene polymers.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of manufacture

If unsubstituted polyfluorene polymers are used, then the structure is simple and easy to manufacture, but the polymers exhibit self-aggregation, loose belt effect, and reduced fluorescence quantum yield

Engineering Contradiction:
Improveease of manufactureVSAvoidfluorescence quantum yield
Core Design Contradiction:
Ease of manufactureVSReliability

Solution Approach 1:

The patent introduces specific substituents at defined positions on the fluorene backbone to create localized structural modifications. The fluoreno[4,5-cde]oxepine unit is incorporated at specific positions along the polymer chain, creating local structural variations that prevent self-aggregation and eliminate the loose belt effect while maintaining overall polymer processability and fluorescence performance

Inventive Principle:
Principle #3Local quality

Solution Approach 2:

The patent creates a composite polymer structure by combining fluorene backbone units with fluoreno[4,5-cde]oxepine substituent units. This composite approach integrates the beneficial properties of both structural elements: the fluorene provides the conjugated backbone for fluorescence, while the fluoreno[4,5-cde]oxepine units provide structural rigidity and prevent aggregation, achieving superior fluorescence quantum yield

Inventive Principle:
Principle #40Composite materials

2Measurement precision

If fluorescent polymers are conjugated to biological molecules, then the detection capability is improved, but the quantum efficiency and function of the fluorescent polymer may be destroyed or reduced

Engineering Contradiction:
Improvedetection capabilityVSAvoidquantum efficiency
Core Design Contradiction:
Measurement precisionVSReliability

Solution Approach 1:

The patent modifies the chemical and physical parameters of the fluorescent polymer through the introduction of fluoreno[4,5-cde]oxepine units, which change the electronic structure and steric properties of the polymer. This parameter change results in enhanced photostability and maintained quantum efficiency even after conjugation to biological molecules, as the modified structure is more resistant to environmental quenching effects

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The fluoreno[4,5-cde]oxepine substituent acts as a protective element that pre-shields the fluorescent backbone from harmful interactions during conjugation with biological molecules. The rigid structure provides steric protection and reduces direct contact between the fluorescent core and quenching biological environments, preserving quantum efficiency before and during the conjugation process

Inventive Principle:
Principle #11Beforehand cushioning (Prior cushioning)

3Reliability

If phycobiliproteins are used as fluorophores, then the extinction coefficient and quantum yield are high, but the proteins are unstable and fade quickly upon illumination

Engineering Contradiction:
Improvequantum yieldVSAvoidphotostability
Core Design Contradiction:
ReliabilityVSDuration of action of stationary object

Solution Approach 1:

The patent replaces the use of unstable phycobiliprotein fluorophores with a synthetic polymer fluorophore that, while having different initial properties, provides superior long-term stability. The fluoreno[4,5-cde]oxepine-containing polymer is designed to be more photostable than natural proteins, eliminating the fading problem while maintaining high quantum yield through its extended conjugated system

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent fundamentally changes the chemical composition and structural parameters from protein-based fluorophores to synthetic polymer fluorophores containing fluoreno[4,5-cde]oxepine units. This parameter change transitions the system from biologically derived, illumination-sensitive proteins to chemically stable, photore resistant synthetic structures that maintain their fluorescence properties over extended illumination periods

Inventive Principle:
Principle #35Parameter changes

4Illumination intensity

If existing polyfluorene polymers are used, then the emission wavelength is close to the UV edge, but this limits the emission wavelength window and increases autofluorescence interference

Engineering Contradiction:
Improveemission wavelength windowVSAvoidautofluorescence interference
Core Design Contradiction:
Illumination intensityVSObject-affected harmful factors

Solution Approach 1:

The patent changes the optical parameters of the polymer by incorporating fluoreno[4,5-cde]oxepine units, which extend the conjugated system and modify the HOMO-LUMO energy gap. This parameter change results in red-shifted emission wavelengths that move away from the UV edge into regions with less biological autofluorescence, expanding the available emission wavelength window for multiplexed detection applications

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent extends the emission spectrum into a new wavelength dimension by incorporating the fluoreno[4,5-cde]oxepine structural motif. This dimensional extension of the emission window allows access to wavelength regions previously unavailable with standard polyfluorene, enabling detection in spectral regions with reduced background interference

Inventive Principle:
Principle #17Another dimension (Dimensionality change)

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

These polymers provide enhanced sensitivity and stability for biological applications, allowing for more effective detection and analysis of analytes with improved fluorescence characteristics and compatibility in aqueous environments.

Implementation Method 1

Fluorescent probes are valuable reagents for the analysis and separation of molecules and cells and for the detection and quantification of other materials

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Implementation Method 2

efficient fluorescence resonance energy transfer

Methodology Applied
Scientific EffectFluorescence resonance energy transfer:

Implementation Method 3

there is a need for fluorescent substances that have strong absorption at 405 nm, and emit fluorescence with a large Stokes shift

Methodology Applied
Scientific EffectLight absorption: Absorption (EM radiation)

Data Source

PatentEP3341377B1Polyfluoreno[4,5-cde]oxepine conjugates and their use in methods of analyte detection
Publication Date: 2021.07.28 AAT BIOQUEST
  • EP3341377B1 patent drawingFigure 1
  • EP3341377B1 patent drawingFigure 2
  • EP3341377B1 patent drawingFigure 3

AI summary

The invention provides for polyfluoreno[4,5-cde]oxepine conjugates and their use in methods of analyte detection.