Protein Detection Composition Using Cyclodextrin and Cellulose
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Solution Overview
Problem
Existing protein detection reagents face challenges with detergent interference, leading to poor stability and high background staining, especially when using acids like phosphoric acid, which results in low sensitivity and laborious washing procedures.
Innovation Solution
A composition comprising a cellulose derivative and a cyclic oligomer, such as cyclodextrin, is used to separate proteins from detergents, allowing for detergent trapping and subsequent protein detection with improved stability and sensitivity, utilizing an acid like tartaric acid to minimize precipitation and enhance staining contrast.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If Coomassie dyes are used in combination with dextrin to overcome detergent interference, then detergent tolerance is improved, but stability deteriorates and background staining increases due to precipitation
Solution Approach 1:
The patent introduces a cyclodextrin derivative as an intermediary substance that specifically binds to detergents through host-guest complexation. This mediator captures the harmful detergent molecules, preventing them from interfering with the Coomassie dye-protein interaction, while the cellulose derivative serves as another intermediary to stabilize the system and prevent precipitation of the dye-cyclodextrin complex
Solution Approach 2:
The invention combines multiple components into a composite staining reagent system: Coomassie dye, cyclodextrin derivative, cellulose derivative, and buffer. This composite formulation synergistically addresses detergent interference through cyclodextrin binding, while the cellulose derivative provides structural stability and prevents precipitation, achieving both detergent tolerance and system stability simultaneously
2Object-affected harmful factors
If washing steps are performed to remove detergent, then detergent interference is reduced, but procedure complexity and time increase
Solution Approach 1:
Instead of removing the detergent through multiple washing steps, the invention converts the harmful detergent presence into a beneficial situation by using cyclodextrin derivatives that specifically bind to detergent molecules. This host-guest complexation traps the detergent in a non-interfering form, allowing direct staining without washing steps while actually improving the detection system's performance
3Measurement precision
If phosphoric acid is used in the staining reagent, then protein detection is enabled, but background staining increases due to precipitation
Solution Approach 1:
The cellulose derivative acts as a protective intermediary that interacts with the phosphoric acid and dye-cyclodextrin complex to prevent unwanted precipitation. This intermediary stabilizes the acidic environment needed for protein staining while preventing the formation of precipitates that would cause background staining, thus maintaining measurement precision without generating harmful background signals
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 solution enables efficient separation of proteins from detergents, reducing background noise and allowing for sensitive protein detection at low levels without the need for extensive washing, using a detergent-tolerant staining system that improves protein detection sensitivity and stability.
Implementation Method 1
the detergent complexation by cyclodextrin is of stoichiometric nature
Implementation Method 2
utilizing an acid like tartaric acid to minimize precipitation and enhance staining contrast
Implementation Method 3
Because detergents adversely affect the colour change on binding of Coomassie dyes to protein
Data Source
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AI summary
The present invention relates to compositions for detection of proteins comprising a protein-complexing dye, a cyclic oligomer, and a hydroxycarboxylic acid, and to methods of detecting and/or quantifying protein comprising contacting a protein-containing sample with such compositions and detecting and/or quantifying the formation of a dye/protein complex.