Protein Target Complex for Selective Dead Cell Labeling
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Solution Overview
Problem
Existing methods for labeling and targeting dead cells are either toxic or unstable, and amine-reactive dyes non-specifically bind to amine groups in the medium, making them unsuitable for in vivo applications.
Innovation Solution
A method using non-activated cyanine and rhodamine compounds that do not cross healthy cell membranes but interact with intracellular proteins in dead cells, allowing for selective labeling and detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If amine-reactive dyes are used to label dead cells, then dead cells can be discriminated from live cells, but the dyes non-specifically bind to amine groups in the medium and are unstable
Solution Approach 1:
The patent changes the chemical parameters of the labeling compounds by using cyanine and rhodamine dyes with specific structural modifications (such as carboxylate groups instead of amine-reactive groups). These parameter changes make the dyes non-activated and prevent non-specific binding to amine groups in the medium, while maintaining their ability to label dead cells through membrane permeability differences.
2Measurement precision
If amine-reactive dyes are used to label dead cells, then dead cells can be discriminated from live cells, but the dyes react with amines in serum components and extracellular proteins
Solution Approach 1:
The patent modifies the chemical parameters of the dye molecules by replacing amine-reactive functional groups with non-activated carboxylate groups. This parameter change eliminates the harmful non-specific binding to serum components and extracellular proteins while preserving the dyes' ability to selectively label dead cells through their cell membrane impermeability properties.
3Measurement precision
If cell-impermeant fluorescent compounds are used to stain nucleic acids, then dead cells can be detected, but the method is toxic and mutagenic
Solution Approach 1:
The patent extracts the toxic and mutagenic properties from the labeling method by abandoning nucleic acid staining entirely. Instead, the invention uses fluorescent compounds that label intracellular proteins in dead cells, thereby achieving dead cell detection without the harmful effects associated with DNA intercalation and staining.
Solution Approach 2:
The patent employs fluorescent compounds (cyanine and rhodamine dyes) that are non-toxic and can be used in physiological conditions. These compounds serve as safe alternatives to mutagenic nucleic acid stains, enabling repeated measurements and in vivo applications without causing cellular damage or mutation.
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 method provides a safe and stable means to discriminate between live and dead cells, enabling in vivo applications and monitoring therapeutic effects.
Implementation Method 1
non-activated cyanine and rhodamine compounds that do not cross healthy cell membranes but interact with intracellular proteins in dead cells
Data Source
AI summary
The present invention relates to a method for labeling or targeting cells whose plasma membrane has lost integrity, such as dead cells, such as for discriminating between live cells and cells whose plasma membrane has lost integrity; to an embodiment of the method for determining one or more values of one or more parameters of cells of a biological sample; to use of the method in an assay for screening drugs for therapy such as cancer therapy; to use of the method to monitor and/or determine the effectiveness of a therapy; to an assay kit; to a complex; to the complex for use as a medicament; to the complex for use in treatment of cancer(s) and/or plaque(s) and/or regeneration and/or supporting the immune system; and, to a dead cell.