CIS Reactive Oxygen Quenchers in Linkers

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

In illuminated reactions, particularly those involving fluorescent or chemiluminescent reactants, prolonged exposure to light sources can cause photodamage, leading to reduced reactant effectiveness and limited signal generation, especially in small reactant volumes used in high-throughput applications like microarrays and single molecule analyses.

Innovation Solution

The development of compounds comprising a nucleoside polyphosphate linked with a photoprotective agent and a dye, where the photoprotective agent is integrated into the structure to mitigate photodamage by reducing the impact of illumination on reactants, thereby extending the photodamage threshold period and maintaining reaction efficiency.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If optically detectable labeling groups (fluorescent or chemiluminescent) are used to monitor reactions, then signal detection capability is improved, but photodamage to reactants increases

Engineering Contradiction:

Improvesignal detection capabilityVSAvoidphotodamage to reactants

Core Design Contradiction:

Measurement precisionVSObject-affected harmful factors

Solution Approach 1:

The patent introduces an oxygen scavenger as an intermediary substance that mediates between the excited fluorescent label and the reactants. The oxygen scavenger consumes ground-state oxygen that would otherwise be converted to reactive singlet oxygen by excited fluorescent labels, thereby protecting reactants from photodamage while allowing signal detection to proceed

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent converts the harmful effect of ground-state oxygen (which is normally present and inert) into a beneficial protective mechanism by introducing oxygen scavengers. The scavengers selectively consume ground-state oxygen, preventing its conversion to harmful singlet oxygen by excited fluorescent labels, thus transforming a potential harm into a protective function

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

2Productivity

If smaller reactant volumes are used in microfluidic or nanofluidic systems, then high throughput and efficiency are improved, but photodamage impact increases and signal generation is limited

Engineering Contradiction:

Improvehigh throughput efficiencyVSAvoidphotodamage impact

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent applies local quality by selectively targeting the local chemical environment within small reactant volumes. Oxygen scavengers are introduced to specifically modify the local oxygen concentration and reactivity in the microfluidic or nanofluidic system, protecting reactants from photodamage at the local level where illumination occurs, while maintaining the overall small volume configuration for high throughput

Inventive Principle:

Principle #3Local quality

3Measurement precision

If prolonged illumination is applied to generate sufficient signal, then detection sensitivity is improved, but reactant damage increases and reaction effectiveness decreases

Engineering Contradiction:

Improvedetection sensitivityVSAvoidreaction effectiveness

Core Design Contradiction:

Measurement precisionVSReliability

Solution Approach 1:

The patent applies preliminary action by pre-introducing oxygen scavengers into the reaction system before illumination begins. These scavengers are already in place to consume ground-state oxygen and prevent its conversion to reactive singlet oxygen during prolonged illumination, thereby protecting reactants and maintaining reaction effectiveness throughout the extended detection period

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent enables continuous useful action by allowing prolonged illumination without significant reactant degradation. Oxygen scavengers continuously consume ground-state oxygen throughout the illumination period, maintaining a protective environment that sustains both signal generation and reactant integrity over extended times, thus achieving continuous detection without loss of reaction effectiveness

Inventive Principle:

Principle #20Continuity of useful action

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 significantly reduces photodamage, allowing reactions to proceed with minimal loss of reactivity, extending the useful time of reactants under illumination and enhancing signal generation in small volumes, thus improving the reliability and accuracy of analyses.

Implementation Method 1

CIS reactive oxygen quenchers integrated into linkers... the photoprotective agent is integrated into the structure to mitigate photodamage by reducing the impact of illumination on reactants

Methodology Applied

Scientific EffectQuenching:

Data Source

PatentUS9200311B2CIS reactive oxygen quenchers integrated into linkers

Publication Date: 2015.12.01 PACIFIC BIOSCIENCES OF CALIFORNIA INC

AI summary

The present invention provides methods and compositions for performing illuminated reactions, particularly sequencing reactions, while mitigating and/or preventing photodamage to reactants that can result from prolonged illumination. In particular, the invention provides methods and compositions for incorporating photoprotective agents into conjugates comprising reporter molecules and nucleoside polyphosphates.