Self-Assembling Fusion Protein Tags for Fast Recombinant Purification
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Solution Overview
Problem
Conventional recombinant protein purification methods are time-consuming, complex, and costly due to the use of expensive beads or resins, requiring multistep processes and are inefficient in achieving high purity.
Innovation Solution
A fusion protein comprising an alpha-helix peptide tag with charged and hydrophobic amino acids forms self-assemblies under specific inducer or condition treatment, allowing purification through simple methods like centrifugation or filtration, eliminating the need for expensive beads or resins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional affinity chromatography using His tag and Ni2+-NTA beads is used for purification, then purification can be achieved, but the process becomes time-consuming and complex with multiple steps
Solution Approach 1:
The invention changes the physical-chemical parameters of the purification system by using a His tag-HAT tag fusion protein that forms reversible self-assemblies under specific conditions (presence/absence of imidazole), enabling purification through simple phase separation rather than complex chromatography steps
Solution Approach 2:
The fusion protein performs self-service by automatically forming self-assemblies that can be easily separated from solution through centrifugation or filtration, eliminating the need for external beads or resins and simplifying the purification process
2Reliability
If conventional affinity chromatography with beads or resins is used, then purification is achieved, but expensive materials and equipment are required
Solution Approach 1:
The invention replaces expensive, reusable beads and resins with a disposable fusion protein tag system that forms transient self-assemblies. The fusion protein itself serves as the purification tool, eliminating the need for costly chromatography media
Solution Approach 2:
The invention extracts the essential purification function from the expensive bead/resin system and transfers it to the fusion protein tag, which forms self-assemblies that can be easily separated and discarded after a single use
3Reliability
If conventional multistep purification process is used, then purification is achieved, but the process complexity increases
Solution Approach 1:
The invention segments the purification process into simple physical separation steps (centrifugation or filtration) based on the formation of self-assemblies, replacing complex chromatography procedures with straightforward phase separation operations
4Manufacturing precision
If conventional affinity chromatography is used, then purification is achieved, but purification purity is lowered due to non-specific binding
Solution Approach 1:
The invention introduces dynamic control of self-assembly formation through imidazole concentration. The fusion protein forms self-assemblies only under specific conditions (low imidazole), allowing dynamic control of purification specificity and reducing non-specific binding
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, efficient, and high-purity purification of recombinant proteins with a simplified process, reducing costs and time, and allows for easy conversion of purified proteins to monomers for further processing.
Implementation Method 1
A fusion protein comprising an alpha-helix peptide tag with charged and hydrophobic amino acids forms self-assemblies under specific inducer or condition treatment
Implementation Method 2
alpha-helix peptide tag with charged and hydrophobic amino acids
Implementation Method 3
alpha-helix peptide tag with charged and hydrophobic amino acids
Implementation Method 4
purification through simple methods like centrifugation or filtration
Implementation Method 5
purification through simple methods like centrifugation or filtration
Data Source
AI summary
The present invention relates to a fusion polypeptide and a method for purifying a target protein by using same, the polypeptide having any one from among a His tag, a HAT tag and an HQ tag, and an α-helix peptide tag of which the major amino acids are charged and hydrophobic amino acids. Since the fusion polypeptide has an ability to induce self-assembly by being fused with the target protein, the target protein purification method using same enables the isolation and purification of the target protein merely through very simple processes such as centrifugation or filtration.