DNA Aptamer Protein Purification Specificity
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Solution Overview
Problem
Current methods for purifying and detecting proteins of interest are inefficient when dealing with complex solutions containing homologous proteins, leading to challenges in achieving high specificity and safety, especially in therapeutic protein production where cross-reactivity can cause adverse immune responses.
Innovation Solution
A method using DNA aptamers immobilized on a solid support via a spacer, which specifically bind to target proteins without binding to homologous proteins, allowing for selective purification and detection by forming complexes that can be recovered or detected.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If antibodies are used for protein purification or detection, then high specificity with respect to target proteins is achieved, but cross-reactivity with homologous proteins occurs leading to reduced purity and potential adverse immune responses
Solution Approach 1:
The patent changes the fundamental binding parameter from antibody-protein interaction to aptamer-protein interaction. DNA aptamers are selected through SELEX to have high specificity for the target protein while showing minimal cross-reactivity with homologous proteins, thereby resolving the contradiction between achieving high specificity and avoiding cross-reactivity
Solution Approach 2:
The patent uses DNA aptamers as synthetic copies that can replicate the specific binding function of antibodies without the immunogenicity and cross-reactivity issues. The aptamers are in vitro selected to mimic antibody specificity while avoiding the problems associated with biological antibody production
2Productivity
If standard purification methods are used for recombinant proteins in transgenic organisms, then production efficiency is maintained, but co-extraction of endogenous homologous proteins occurs reducing product safety
Solution Approach 1:
The patent extracts only the desired recombinant protein from the complex mixture by using aptamers that specifically bind to the transgenic protein sequence while not binding to the endogenous homologous protein. This selective extraction removes the contaminating homologous proteins while maintaining high production efficiency
Solution Approach 2:
The patent introduces DNA aptamers as intermediary binding agents that mediate the selective separation of recombinant proteins from endogenous homologous proteins. The aptamers serve as a bridge between the target protein and the purification system, enabling specific isolation without affecting productivity
3Object-affected harmful factors
If high specificity purification is implemented to eliminate homologous proteins, then product safety is improved, but purification complexity and cost increase
Solution Approach 1:
The patent uses DNA aptamers which are chemically synthesized and can be produced cheaply and rapidly compared to antibodies. The aptamers can be easily replaced and do not require complex biological production systems, thereby reducing purification complexity while maintaining high product safety
Solution Approach 2:
The patent replaces the complex biological system of antibody production and purification with a simpler chemical synthesis and in vitro selection system for aptamers. This substitution eliminates the need for animal immunization, hybridoma culture, and antibody purification steps, reducing overall system complexity while achieving high specificity
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 enhances the specificity and safety of protein purification and detection, reducing the risk of adverse immune responses and improving the harmlessness of therapeutic proteins by effectively discriminating between target and homologous proteins, even in the presence of small quantities of homologous proteins.
Implementation Method 1
a DNA aptamer which specifically binds to a determined target protein
Data Source
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AI summary
The invention relates to a method for purifying or detecting a target protein present in a solution, wherein said method comprises, before carrying out the purification or detection step itself, the step of contacting said solution with an aptamer binding specifically to said target protein, wherein said aptamer does not bind to any protein homologous to said target protein that could also be present in the solution.