Fusion Tag Fluorophore Detection in Capillary Electrophoresis
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Solution Overview
Problem
Current methods for detecting fusion proteins in high throughput settings are either not suitable for complex samples like cell lysates or are non-qualitative, limiting the ability to rapidly characterize starting materials in protein production pipelines.
Innovation Solution
The use of fusion tag-specific fluorescent labels coupled to proteins of interest, followed by automated capillary electrophoresis, allows for high throughput detection and purification by specifically binding to the fusion tags and generating fluorescence signals for protein detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If western blot methods are used to detect fusion proteins, then specific detection capability is achieved, but throughput is low and processing time is long
Solution Approach 1:
The patent extracts the detection function from complex western blot procedures by using fusion tag-specific fluorescent labels that directly bind to the fusion tag, eliminating the need for antibodies and extensive sample preparation while maintaining specific detection capability
Solution Approach 2:
The patent replaces the mechanical transfer and blotting steps of western blot with fluorescence-based detection in capillary electrophoresis, substituting a complex mechanical process with a simpler optical detection method that achieves both specificity and high throughput
2Productivity
If ELISA methods are used for protein detection, then high throughput is achieved, but detection results are non-qualitative
Solution Approach 1:
The patent uses fluorescence intensity variations to provide both quantitative and qualitative detection information, where the presence, absence, and intensity of fluorescence signals indicate protein expression levels and characteristics, transforming non-qualitative ELISA results into informative qualitative data
3Productivity
If capillary electrophoresis is used for protein analysis, then high throughput and rapid processing are achieved, but specific detection capability in complex samples is lost
Solution Approach 1:
The patent introduces fusion tag-specific fluorescent labels as intermediaries that specifically bind to the fusion tag on proteins of interest, enabling selective detection of target proteins within complex samples like cell lysates while maintaining the high throughput capability of capillary electrophoresis
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 enables rapid, qualitative, and quantitative detection of fusion proteins in complex samples, improving the efficiency of protein production pipelines by providing a high throughput method for protein characterization without the need for antibodies or extensive sample preparation.
Implementation Method 1
the presence of the proteins of interest in the sample is detected by fluorescence signals associated with the fusion tag specific fluorophores
Implementation Method 2
subjecting the sample to automated capillary electrophoresis
Data Source
AI summary
A method and related microfluidic chip and kit for high throughput detection of proteins of interest contained in a sample is disclosed. The method comprises of specifically labeling fusion proteins in a complex sample with fusion tag specific fluorophores that specifically bind the fusion tags coupled to the proteins of interest, and subjecting the sample to automated capillary electrophoresis, wherein the presence of the proteins of interest in the sample is detected by fluorescence signals associated with the fusion tag specific fluorophores.