Self-Assembling Peptide Tags for Recombinant Protein 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, and require multistep processes, making scale-up difficult and increasing production costs.
Innovation Solution
A peptide tag composed of charged and polar amino acids forms self-assemblies under specific inducer or condition treatment, allowing for simple purification methods like centrifugation or filtration, eliminating the need for expensive beads or resins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional purification methods using His tag and Ni 2+ affinity chromatography are employed, then recombinant proteins can be purified, but the process requires expensive beads or resins, multistep complex procedures, and consumes a lot of time
Solution Approach 1:
The invention changes the physical-chemical parameters of the purification system by using a peptide tag composed of charged and polar amino acids that undergoes condensation polymerization to form macromolecular assemblies. This phase transition from soluble monomers to insoluble macromolecular structures enables simple separation by centrifugation or filtration, replacing complex affinity chromatography procedures
Solution Approach 2:
The invention extracts the purification function from complex chromatography systems to a simple self-assembly system. The peptide tag autonomously forms macromolecular assemblies that can be separated from the solution through simple physical methods, eliminating the need for expensive beads, resins, and complex multistep procedures
2Reliability
If conventional affinity chromatography with Ni 2+ and His tag is used, then protein purification can be achieved, but expensive beads or resins and expensive devices or equipment are required
Solution Approach 1:
The invention replaces expensive, reusable chromatography beads and resins with a disposable peptide tag system. The peptide tag is incorporated into the recombinant protein during expression, and after purification, the tag can be removed by protease cleavage. This eliminates the need for expensive purification materials while maintaining effective purification
Solution Approach 2:
The peptide tag autonomously performs the purification function through self-assembly into macromolecular structures. The tag does not require external affinity ligands or complex equipment; it spontaneously forms separable structures under specific conditions, making the purification system self-sufficient and eliminating dependency on expensive external reagents
3Reliability
If conventional multistep purification processes are employed, then high purity recombinant proteins can be obtained, but the process consumes a lot of time and is difficult to scale up
Solution Approach 1:
The invention segments the purification process into two simple steps: (1) induction of macromolecular assembly formation by adding specific ions or adjusting pH, and (2) separation by centrifugation or filtration. This segmentation replaces complex multistep procedures with simple, rapid operations that can be easily scaled up for large-volume production
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 reduced costs by inducing macromolecular self-assembly, facilitating easy conversion to monomers and tag removal.
Implementation Method 1
peptide tag having main amino acids composed of charged and polar amino acids and forming self-assembly
Implementation Method 2
charged and polar amino acids forming self-assembly
Implementation Method 3
charged and polar amino acids forming self-assembly
Implementation Method 4
purification methods like centrifugation or filtration
Implementation Method 5
purification methods like centrifugation or filtration
Data Source
Figure 1a
Figure 1b
Figure 1c
AI summary
The present invention relates to: a fusion polypeptide in which a His tag is fused with a peptide tag having main amino acids composed of charged and polar amino acids; a fusion protein in which the fusion polypeptide is fused with a target protein; and a target protein purification method using the same. Since the peptide tag is fused with the target protein and thus has the ability to induce the formation of a self-assembly, a target protein purification method using same can be used to isolate and purify the target protein by using only a very simple method such as centrifugation or filtration.